Analysis of Variation in Clubfoot Candidate Genes

Isolated clubfoot, a common birth defect occurring in more than 135,000 livebirths worldwide each year, is associated with significant health care and financial burdens. Clubfoot is defined by forefoot adduction, hindfoot varus, midfoot cavus and hindfoot equinus. Isolated clubfoot, which is the focus of these studies, is distinct from syndromic clubfoot because there are no other associated malformations. Population, family, twin and segregation analysis studies provide evidence that genetic and environmental factors play an etiologic role in isolated clubfoot. The studies described in this thesis were performed to define the role of genetic variation in isolated clubfoot. Interrogation of a deletion region associated with syndromic clubfoot, suggested that CASP8 and CASP10, two apoptotic genes, play a role in isolated clubfoot. To explore the role of apoptotic genes in clubfoot, SNPs spanning genes involved in the apoptotic pathway in the six chromosomal deletion regions, and limb patterning genes, HOXD and HOXA, were interrogated. SNPs in mitochondrial mediated apoptotic genes and several SNPs in HOXA and HOXD genes were modestly associated with clubfoot with the most significant SNP, rs3801776, located in the basal promoter of HOXA9. Several significant associations were found with SNPs in NFAT2 and TNIP2. Significant gene interactions were detected between SNPs in HOX and apoptotic genes. These findings suggest a model for clubfoot in which variation in one gene is not sufficient to cause the malformation but requires variation several genes to perturb protein expression sufficiently to alter muscle and foot development. These results significantly impact our knowledge base by delineating underlying mechanisms causing clubfoot.

Assembling Multiporphyrin Stacks Inside the DNA Double Helix

Double stranded DNA hybrids containing up to four consecutive, face-to-face stacked porphyrins are described. Non-nucleosidic, 5,15-bisphenyl-substituted porphyrin building blocks were incorporated into complementary oligonucleotide strands. Upon hybridization multiple porphyrins are well accommodated inside the DNA scaffold without disturbing the overall B-DNA structure. The formation of double strands containing up to four free base porphyrins is enabled without compromising duplex stability. UV/vis, fluorescence, and CD spectroscopy demonstrate the formation of porphyrins H-aggregates inside the DNA double helix and provide evidence for the existence of strong excitonic coupling between interstrand stacked porphyrins. H-aggregation results in considerable fluorescence quenching. Most intense CD effects are observed in stacks containing four porphyrins. The findings demonstrate the value of DNA for the controlled formation of molecularly defined porphyrin aggregates.

Remote Effects Modulating the Spin Equilibrium of the Resting State of Cytochrome P450cam –An Investigation Using Active Site Analogues

The crystal structure of the resting state of cytochrome P450cam (CYP101), a heme thiolate protein, shows a cluster of six water molecules in the substrate binding pocket, one of which is coordinating to iron(III) as sixth ligand. The resting state is low-spin and changes to high-spin when substrate camphor binds and H2O is removed. In contrast to the protein, previously synthesised enzyme models such as H2O[BOND]FeIII(porph)(ArS−) were shown to be purely high-spin. Iron(S−)porphyrins with different distal sites mimicking proposed remote effects have been prepared and studied by cw-EPR. The results indicate that the low-spin of the resting state of P450cam is due to the fact that the water molecule coordinating to iron has an OH−-like character because of hydrogen bonding and polarisation of the water cluster, respectively.

The origin of the low-spin character of the resting state of cytochrome P450cam investigated by means of active site analogues

Crown-capped iron(S−) porphyrins 1·H2O and 2·H2O and their corresponding Ba2+ complexes have been prepared as active site analogues of the resting state of cytochrome P450cam. cw-EPR studies and electronic structure calculations at the density functional theory (DFT) level of model systems suggest a functional role of the water cluster of P450cam.

SNPs for parentage testing and traceability in globally diverse breeds of sheep

DNA-based parentage determination accelerates genetic improvement in sheep by increasing pedigree accuracy. Single nucleotide polymorphism (SNP) markers can be used for determining parentage and to provide unique molecular identifiers for tracing sheep products to their source. However, the utility of a particular "parentage SNP" varies by breed depending on its minor allele frequency (MAF) and its sequence context. Our aims were to identify parentage SNPs with exceptional qualities for use in globally diverse breeds and to develop a subset for use in North American sheep. Starting with genotypes from 2,915 sheep and 74 breed groups provided by the International Sheep Genomics Consortium (ISGC), we analyzed 47,693 autosomal SNPs by multiple criteria and selected 163 with desirable properties for parentage testing. On average, each of the 163 SNPs was highly informative (MAF≥0.3) in 48±5 breed groups. Nearby polymorphisms that could otherwise confound genetic testing were identified by whole genome and Sanger sequencing of 166 sheep from 54 breed groups. A genetic test with 109 of the 163 parentage SNPs was developed for matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The scoring rates and accuracies for these 109 SNPs were greater than 99% in a panel of North American sheep. In a blinded set of 96 families (sire, dam, and non-identical twin lambs), each parent of every lamb was identified without using the other parent's genotype. In 74 ISGC breed groups, the median estimates for probability of a coincidental match between two animals (PI), and the fraction of potential adults excluded from parentage (PE) were 1.1×10(-39) and 0.999987, respectively, for the 109 SNPs combined. The availability of a well-characterized set of 163 parentage SNPs facilitates the development of high-throughput genetic technologies for implementing accurate and economical parentage testing and traceability in many of the world's sheep breeds.

Thermal and collisional history of Tishomingo iron meteorite: More evidence for early disruption of differentiated planetesimals

Tishomingo is a chemically and structurally unique iron with 32.5 wt.% Ni that contains 20% residual taenite and 80% martensite plates, which formed on cooling to between -75 and -200 °C, probably the lowest temperature recorded by any meteorite. Our studies using transmission (TEM) and scanning electron microscopy (SEM), X-ray microanalysis (AEM) and electron backscatter diffraction (EBSD) show that martensite plates in Tishomingo formed in a single crystal of taenite and decomposed during reheating forming 10-100 nm taenite particles with ∼50 wt.% Ni, kamacite with ∼4 wt.%Ni, along with martensite or taenite with 32 wt.% Ni. EBSD data and experimental constraints show that Tishomingo was reheated to 320-400 °C for about a year transforming some martensite to kamacite and to taenite particles and some martensite directly to taenite without composition change. Fizzy-textured intergrowths of troilite, kamacite with 2.7 wt.% Ni and 2.6 wt.% Co, and taenite with 56 wt.% Ni and 0.15 wt.% Co formed by localized shock melting. A single impact probably melted the sub-mm sulfides, formed stishovite, and reheated and decomposed the martensite plates. Tishomingo and its near-twin Willow Grove, which has 28 wt.% Ni, differ from IAB-related irons like Santa Catharina and San Cristobal that contain 25-36 wt.% Ni, as they are highly depleted in moderately volatile siderophiles and enriched in Ir and other refractory elements. Tishomingo and Willow Grove therefore resemble IVB irons but are chemically distinct. The absence of cloudy taenite in these two irons shows that they cooled through 250 °C abnormally fast at >0.01 °C/yr. Thus this grouplet, like the IVA and IVB irons, suffered an early impact that disrupted their parent body when it was still hot. Our noble gas data show that Tishomingo was excavated from its parent body about 100 to 200 Myr ago and exposed to cosmic rays as a meteoroid with a radius of ∼50-85 cm.

Post-Empire Imaginaries? Anglophone Literature, History, and the Demise of Empires

Empires as political entities may be a thing of the past, but as a concept, empire is alive and kicking. From heritage tourism and costume dramas to theories of the imperial idea(l): empire sells. Post-Empire Imaginaries? Anglophone Literature, History, and the Demise of Empires presents innovative scholarship on the lives and legacies of empires in diverse media such as literature, film, advertising, and the visual arts. Though rooted in real space and history, the post-empire and its twin, the post-imperial, emerge as ungraspable ideational constructs. The volume convincingly establishes empire as welcoming resistance and affirmation, introducing post-empire imaginaries as figurations that connect the archives and repertoires of colonial nostalgia, postcolonial critique, post-imperial dreaming.

Placental plasticity in monochorionic twins: Impact on birth weight and placental weight

INTRODUCTION The knowledge about adaptive mechanisms of monochorionic placentas to fulfill the demands of two instead of one fetus is largely speculative. The aim of our study was to investigate the impact of chorionicity on birth weight and placental weight in twin pregnancies. METHODS Forty Monochorionic (MC) and 43 dichorionic (DC) twin pregnancies were included in this retrospective study. Individual and total (sum of both twins) birth weights, placental weights ratios between placental and birth weights and observed-to-expected (O/E)-ratios were calculated and analyzed. Additionally, we investigated whether in twin pregnancies placental and birth weights follow the law of allometric metabolic scaling. RESULTS MC pregnancies showed higher placental O/E-ratios than DC ones (2.25 ± 0.85 versus 1.66 ± 0.61; p < 0.05), whereas the total neonatal birth weight O/E-ratios were not different. In DC twins total placental weights correlated significantly with gestational age (r = 0.74, p < 0.001), but not in MC twins. Analysis of deliveries ≤32 weeks revealed that the placenta to birth weight ratio in MC twins was higher than in matched DC twins (0.49 ± 0.3 versus 0.24 ± 0.03; p = 0.03). Allometric metabolic scaling revealed that dichorionic twin placentas scale with birth weight, while the monochorionic ones do not. DISCUSSION The weight of MC placentas compared to that of DC is not gestational age dependent in the third trimester. Therefore an early accelerated placental growth pattern has to be postulated which leads to an excess placental mass particularly below 32 weeks of gestation. The monochorionic twins do not follow allometric metabolic scaling principle making them more vulnerable to placental compromise.

Knockdown of Kiaa0319 Reduces Dendritic Spine Density

Developmental Dyslexia is a reading disorder that affects individuals that possess otherwise normal intelligence. Until the four candidate dyslexia susceptibility genes were discovered, the cause of cortical malformations found in post mortem dyslexic brains was unclear. Normal brain development is crucial for the proper wiring of the neural circuitry that allow an individual to perform cognitive tasks like reading. For years, familial and twin studies have suggested that there was a genetic basis to the causation of dyslexia. Kiaa0319 was among the candidate dyslexia susceptibility genes that were ascertained. KIAA0319 is located on Chromosome 6p22.2-22.3 and has been found to exhibit differential spatial-temporal expression patterns in the brain throughout development, which suggests that the polycystic kidney disease (PKD) domain encoded by KIAA0319 facilitates cell-cell adhesion to enable neuronal precursors to crawl up the radial glia during neuronal migration. With the knowledge of KIAA0319 involvement in early neurogenesis, we were interested in determining how different KIAA0319 expression may impact cortical neurons in layer II and III during early adulthood. We show that KIAA0319 knockdown in cortical pyramidal neurons significantly reduces the dendritic spine density. Studies have shown that changes in dendritic spine morphology and density affect properties of neural circuitry. Henceforth, this finding may reveal a link between the Kiaa0319 gene and the deficit of the neural processing task of reading due to reduced spines density. Finding a correlation between Kiaa0319 expression and its influence on dendritic spine development may lead to a greater insight of a direct link between the dyslexia susceptibility gene and the biological mechanism that causes dyslexia.

A case-control study of the risk factors for eclampsia

Objective. To study the risk factors for eclampsia, a rare but significant complication of pregnancy.^ Target population. All deliveries at or after the 20th week of gestation that took place between January 1, 1977 and March 1992, and between January 1990 and April 1992 at two hospitals in Houston, Texas, respectively.^ Study population. Sixty-six confirmed cases of eclampsia, and 2 groups of randomly selected controls: Non-preeclamptic and preeclamptic deliveries matched to cases on hospital and month of delivery on a 1:4 ratio.^ Exclusions. Women with chronic hypertension, gestational epilepsy, a previous history of epilepsy, and convulsions attributed to encephalitis, meningitis, cerebral tumor, and intracerebral bleeding, and women without a definite diagnosis of preeclampsia/eclampsia.^ Results. Eclampsia developed in 0.52-0.93/1000 deliveries. Fifty-six percent of seizures occurred in the antepartum period, 2% as early as 20 weeks of gestation and 39% between 37 and 42 weeks. Twenty-nine percent and 15% occurred in the postpartum and late postpartum periods, respectively, 8% as late as one week postpartum. A different set of risk factors was involved in the development of eclampsia in non-preeclamptic women than in the progression from preeclampsia to eclampsia. Factors involved in the development of eclampsia included, in addition to twin pregnancy and family history of pregnancy-induced hypertension, fewer than 3 prenatal care visits, urinary tract infections, primigravidity, obesity, black ethnicity, diabetes mellitus, and age $\le$20 years. Risk factors involved in the progression from preeclampsia to eclampsia included fewer than 3 of prenatal care visits, and age $\le$20 years. Protective factors were magnesium sulfate administration prior to seizure, history of abortions and longer gestational age. Having less than 3 prenatal care visits and being less than or equal to 20 years of age were predictors of eclampsia, whether of its development or progression from preeclampsia. Once preeclampsia is diagnosed, primigravid, diabetic, black, or obese women and those with urinary tract infections did not appear to exhibit any increased risk for the progression to eclampsia. The administration of magnesium sulfate was especially protective, followed by a positive history of abortions, 3 or more prenatal care visits, and longer gestational age. The protective effect of MgSO$\sb4$ was only slightly diminished when cases were restricted to the 65% who had a diagnosis of preeclampsia. The progression from preeclampsia to eclampsia may be largely preventable through adequate prenatal care and presumably the administration of magnesium sulfate. (Abstract shortened by UMI.) ^

Evaluation of Narrow Row Soybeans

Producers continue to look at different management practices to increase corn and soybean yields. One area of interest is planting corn and soybeans in narrow rows. Traditionally, these crops have been planted in row widths of 30 to 38 in. Planters on the market today have the capability to plant corn and soybeans in 15- and 20-in. rows, as well as in twin rows that are spaced eight inches apart. This study was set up to evaluate the yield impact of planting soybeans in 15-in. rows versus the traditional 30-in. row spacing.

Chronostratigraphic framework at sites GeoB14403 and GeoB14414 from the Gela Basin, Sicily Channel

A multi-proxy chronological framework along with sequence-stratigraphic interpretations unveils composite Milankovitch cyclicity in the sedimentary records of the Last GlacialeInterglacial cycle at NE Gela Basin on the Sicilian continental margin. Chronostratigraphic data (including foraminifera-based eco-biostratigraphy and d18O records, tephrochronological markers and 14C AMS radiometric datings) was derived from the shallow-shelf drill sites GeoB14403 (54.6 m recovery) and GeoB14414 (27.5 m), collected with both gravity and drilled MeBo cores in 193 m and 146 m water depth, respectively. The recovered intervals record Marine Isotope Stages and Substages (MIS) from MIS 5 to MIS 1, thus comprising major stratigraphic parts of the progradational deposits that form the last 100-ka depositional sequence. Calibration of shelf sedimentary units with borehole stratigraphies indicates the impact of higher-frequency (20-ka) sea level cycles punctuating this 100-ka cycle. This becomes most evident in the alternation of thick interstadial highstand (HST) wedges and thinner glacial forced-regression (FSST) units mirroring seaward shifts in coastal progradation. Albeit their relatively short-lived depositional phase, these subordinate HST units form the bulk of the 100-ka depositional sequence. Two mechanisms are proposed that likely account for enhanced sediment accumulation ratios (SAR) of up to 200 cm/ka during these intervals: (1) intensified activity of deep and intermediate Levantine Intermediate Water (LIW) associated to the drowning of Mediterranean shelves, and (2) amplified sediment flux along the flooded shelf in response to hyperpycnal plumes that generate through extreme precipitation events during overall arid conditions. Equally, the latter mechanism is thought to be at the origin of undulated features resolved in the acoustic records of MIS 5 Interstadials, which bear a striking resemblance to modern equivalents forming on late-Holocene prodeltas of other Mediterranean shallow-shelf settings.