Coherent population accumulations of multiphoton transitions induced by an ultrashort pulse train in polar molecules

Coherent population accumulations of multiphoton transitions induced by an ultrashort pulse train in a two-level polar molecule are investigated theoretically by solving the density-matrix equations without invoking any of the standard approximations. It is shown due to the effects of permanent dipole moments, that the population accumulation of multiphoton transitions can be obtained in the polar molecule. Moreover, the population accumulations depend crucially on the relative phase between two sequential pulses, and the period in which the maximum population accumulation occurs is 2 pi/N in N-photon transitions.

Functional evaluation of noncovalent interactions in neuroreceptors and progress toward the expansion of unnatural amino acid methodology

This dissertation primarily describes chemical-scale studies of G protein-coupled receptors and Cys-loop ligand-gated ion channels to better understand ligand binding interactions and the mechanism of channel activation using recently published crystal structures as a guide. These studies employ the use of unnatural amino acid mutagenesis and electrophysiology to measure subtle changes in receptor function.

In chapter 2, the role of a conserved aromatic microdomain predicted in the D3 dopamine receptor is probed in the closely related D2 and D4 dopamine receptors. This domain was found to act as a structural unit near the ligand binding site that is important for receptor function. The domain consists of several functionally important noncovalent interactions including hydrogen bond, aromatic-aromatic, and sulfur-π interactions that show strong couplings by mutant cycle analysis. We also assign an alternate interpretation for the linear fluorination plot observed at W6.48, a residue previously thought to participate in a cation-π interaction with dopamine.

Chapter 3 outlines attempts to incorporate chemically synthesized and in vitro acylated unnatural amino acids into mammalian cells. While our attempts were not successful, method optimizations and data for nonsense suppression with an in vivo acylated tRNA are included. This chapter is aimed to aid future researchers attempting unnatural amino acid mutagenesis in mammalian cells.

Chapter 4 identifies a cation-π interaction between glutamate and a tyrosine residue on loop C in the GluClβ receptor. Using the recently published crystal structure of the homologous GluClα receptor, other ligand-binding and protein-protein interactions are probed to determine the similarity between this invertebrate receptor and other more distantly related vertebrate Cys-loop receptors. We find that many of the interactions previously observed are conserved in the GluCl receptors, however care must be taken when extrapolating structural data.

Chapter 5 examines inherent properties of the GluClα receptor that are responsible for the observed glutamate insensitivity of the receptor. Chimera synthesis and mutagenesis reveal the C-terminal portion of the M4 helix and the C-terminus as contributing to formation of the decoupled state, where ligand binding is incapable of triggering channel gating. Receptor mutagenesis was unable to identify single residue mismatches or impaired protein-protein interactions within this domain. We conclude that M4 helix structure and/or membrane dynamics are likely the cause of ligand insensitivity in this receptor and that the M4 helix has an role important in the activation process.

Engineered underdominance as a method of insect population replacement and reproductive isolation

Insect vector-borne diseases, such as malaria and dengue fever (both spread by mosquito vectors), continue to significantly impact health worldwide, despite the efforts put forth to eradicate them. Suppression strategies utilizing genetically modified disease-refractory insects have surfaced as an attractive means of disease control, and progress has been made on engineering disease-resistant insect vectors. However, laboratory-engineered disease refractory genes would probably not spread in the wild, and would most likely need to be linked to a gene drive system in order to proliferate in native insect populations. Underdominant systems like translocations and engineered underdominance have been proposed as potential mechanisms for spreading disease refractory genes. Not only do these threshold-dependent systems have certain advantages over other potential gene drive mechanisms, such as localization of gene drive and removability, extreme engineered underdominance can also be used to bring about reproductive isolation, which may be of interest in controlling the spread of GMO crops. Proof-of-principle establishment of such drive mechanisms in a well-understood and studied insect, such as Drosophila melanogaster, is essential before more applied systems can be developed for the less characterized vector species of interest, such as mosquitoes. This work details the development of several distinct types of engineered underdominance and of translocations in Drosophila, including ones capable of bringing about reproductive isolation and population replacement, as a proof of concept study that can inform efforts to construct such systems in insect disease vectors.

Transient population and polarization gratings induced by (1+1)-dimensional ultrashort dipole soliton

An ultrafast transient population grating induced by a (1+1)-dimensional, ultrashort dipole soliton is demonstrated by solving the full-wave Maxwell-Bloch equations. The number of lines and the period of the grating can be controlled by the beam waist and the area of the pulse. Of interest is that a polarization grating is produced. A coherent control scheme based on these phenomena can be contemplated as ultrafast transient grating techniques.

Cheese consumption and prevalence of overweight and obesity in a Basque adult population: a crosssectional study

[EN] Studies have reported a negative association between dairy product consumption and weight status. However, not as much research has focused on cheese; therefore, the aim of this study was to study the association between cheese intake and overweight and obesity in a representative Basque adult population. A food frequency questionnaire (FFQ) was obtained from a random sample of 1081 adults (530 males and 551 females, 17–96 years old). Cheese consumption data were expressed as g/1000 kcal/day. The prevalence of overweight/obesity was higher in men (55.1%) than in women (35.4%) (p50.001). Participants with low or moderate intake of fresh and processed cheese demonstrated a higher prevalence of excess weight, compared with those with higher consumption. The confounding variables selected in multivariate analysis were occupational status and age in both genders; and place of residence in men. In conclusion, negative associations were found between consumption of some types of cheese and overweight and obesity in this population.

Structure-Guided Design and Biophysical Characterization of Novel Anti-HIV Reagents

Despite over 30 years of effort, an HIV-1 vaccine that elicits protective antibodies still does not exist. Recent clinical studies have identified that during natural infection about 20% of the population is capable of mounting a potent and protective antibody response. Closer inspection of these individuals reveal that a subset of these antibodies, recently termed potent VRC01-like (PVL), derive exclusively from a single human germline heavy chain gene. Induced clonal expansion of the B cell encoding this gene is the first step through which PVL antibodies may be elicited. Unfortunately, naturally occurring HIV gp120s fail to bind to this germline, and as a result cannot be used as the initial prime for a vaccine regimen. We have determined the crystal structure of an important germline antibody that is a promising target for vaccine design efforts, and have set out to engineer a more likely candidate using computationally-guided rational design.

In addition to prevention efforts on the side of vaccine design, recently characterized broadly neutralizing anti-HIV antibodies have excellent potential for use in gene therapy and passive immunotherapy. The separation distance between functional Fabs on an antibody is important due to the sparse distribution of envelop spikes on HIV compared to other viruses. We set out to build and characterize novel antibody architectures by incorporating structured linkers into the hinge region of an anti-HIV antibody b12. The goal was to observe whether these linkers increased the arm-span of the IgG dimer. When incorporated, flexible Gly4Ser repeats did not result in detectable extensions of the IgG antigen binding domains, by contrast to linkers including more rigid domains such as β2-microglobulin, Zn-α2-glycoprotein, and tetratricopeptide repeats (TPRs). This study adds an additional set of linkers with varying lengths and rigidities to the available linker repertoire, which may be useful for the modification and construction of antibodies and other fusion proteins.

Morphological variation with interspecific implications in a population of Sigara dorsalis (Leach) (Hemiptera Heteroptera: Corixidae), and a request for British specimens

Sigara dorsalis belongs to a very closely related group of six species forming the sub-genus Sigara sensu strictu. Each of the six species has a distinct allopatric geographical distribution in Europe. Studies were started on a series of populations in the north west Midlands of England. All the populations examined, except one, contained only males with the typical diagnostic features of S. dorsalis, albeit with considerable variation. One pond near Congleton, Cheshire situated in a permanent-ley pasture and apparently free from pollution contained typical S. dorsalis males but, in addition, many atypical individuals. From one sample of forty-six males, all possessed left parameres with the slight point on the dorsal surface characteristic of S. dorsalis. However, almost half possessed additional morphological features intermediate between S. dorsalis and S. striata.

Quantum interference effects of two coherent population trapping states on the atomic spectral lines of a F-e=0 -> F-g=1 transition

We investigate the fluorescence spectrum in a nearly degenerate atomic system of a F-e = 0 -> F-g = 1 transition by analytically solving Schrodinger equations. An ultranarrow fluorescence spectral line in between the two coherent population trapping windows has been found. Our analytic solutions clearly show the origin of the ultranarrow spectral line. Due to quantum interference effects between two coherent population trapping states, the width and intensity of the central spectral line can be controlled by an external magnetic field. Such an effect may be used to detect a magnetic field.

Restoration of a brown trout (Salmo trutta L.) population to Loch Enoch, an acified Loch in Galloway, South-West Scotland

The authors present the findings of a restoration project in Loch Enoch in Scotland. There are historical references that brown trout was present in Loch Enoch up to the 1920s but it is believed the acidity of loch triggered the disappearance of Salmo trutta. The recent observed reduction in the acidity of L. Enoch to a level close to that found in nearby lochs with trout populations, suggested that trout might now survive in L. Enoch. For a population to survive, all stages in the life-cycle of a species must be able to develop. Accordingly, tests were undertaken, first with eggs and fry. The availability of food was also studied. In October 1994, 3,000 yearling trout of L. Grannoch origin which had been reared in a local hatchery were distributed throughout the loch. The fish population was studied from 1995-98. The authors conclude that survival of the trout population is possible if the acidity of the loch water remains low.

Temperature dependence of dark lines in coherent population trapping

We measure the signal amplitude and linewidth of a dark line in coherent population trapping in the Rb vapour cell filled with mixed buffer gas N-2 and Ar as a function of cell temperature. We find that the dark line signal amplitude increases with temperature up to a maximum at 49 degrees C and then drops at higher temperatures due to quenching effects of N-2. The linewidth of the dark line remains basically constant, at 1080 Hz. We also measure the linewidth of the dark line as a function of laser intensity. The linewidth increases linearly with laser intensity. An intrinsic linewidth (FWHM=896 Hz at 3.4 GHz) of the Rb cell is obtained.