Physical Modeling And Parametric Study On Two-Degree-Of-Freedom Viv Of A Cylinder Near Rigid Wall

Unlike most previous studies on the transverse vortex-induced vibration(VIV) of a cylinder mainly under the wallfree condition (Williamson & Govardhan,2004),this paper experimentally investigates the vortex-induced vibration of a cylinder with two degrees of freedom near a rigid wall exposed to steady flow.The amplitude and frequency responses of the cylinder are discussed.The lee wake flow patterns of the cylinder undergoing VIV were visualized by employing the hydrogen bubble technique.The effects of the gap-to-diameter ratio (e0/D) and the mass ratio on the vibration amplitude and frequency are analyzed.Comparisons of VIV response of the cylinder are made between one degree (only transverse) and two degrees of freedom (streamwise and transverse) and those between the present study and previous ones.The experimental observation indicates that there are two types of streamwise vibration,i.e.the first streamwise vibration (FSV) with small amplitude and the second streamwise vibration (SSV) which coexists with transverse vibration.The vortex shedding pattem for the FSV is approximately symmetric and that for the SSV is alternate.The first streamwise vibration tends to disappear with the decrease of e0/D.For the case of large gap-to-diameter ratios (e.g.e0/D = 0.54～1.58),the maximum amplitudes of the second streamwise vibration and transverse one increase with the increasing gapto-diameter ratio.But for the case of small gap-to-diameter ratios (e.g.e0/D = 0.16,0.23),the vibration amplitude of the cylinder increases slowly at the initial stage (i.e.at small reduced velocity V,),and across the maximum amplitude it decreases quickly at the last stage (i.e.at large Vr).Within the range ofthe examined small mass ratio (m＜4),both streamwise and transverse vibration amplitude of the cylinder decrease with the increase of mass ratio for the fixed value of V,.The vibration range (in terms of Vr ) tends to widen with the decrease of the mass ratio.In the second streamwise vibration region,the vibration frequency of the cylinder with a small mass ratio (e.g.mx = 1.44) undergoes a jump at a certain Vr,.The maximum amplitudes of the transverse vibration for two-degree-of-freedom case is larger than that for one-degree-of-freedom case,but the transverse vibration frequency of the cylinder with two degrees of freedom is lower than that with one degree of freedom (transverse).

An overview of the Clupeid fishing gear of Kainji Lake

Fishing gears and methods that target the Clupeids are atalla lift net, light attraction, midwater trawl and dala (Clupeid beach seine).Dala fisheries have been the most lucrative fishing gear on Clupeids within the recent past. However, it has been declared as 100% illegal gear having a mesh size of 0.1 mm, because it catches indiscriminately undersized (juveniles/larvae) commercial fish. The ban on the gear was therefore mounted through promulgation and implementation of special fisheries laws for Kainji Lake (Nigeria). Although, the use of Dala fishing gear and method has decreased due to this management approach, its use is currently increased again. It is therefore suggested that the ban on the use of Dala may not be the answer to indiscriminate fisheries, but modification of the gear and methods for harvesting Clupeids should be the main objective

Noninterferometric measurement of lead zirconate titanate inverse piezoelectric extension

A noncontacting and noninterferometric depth discrimination technique, which is based on differential confocal microscopy, was used to measure the inverse piezoelectric extension of a piezoelectric ceramic lead zirconate titanate actuator. The response characteristics of the actuator with respect to the applied voltage, including displacement, linearity, and hysteresis, were obtained with nanometer measurement accuracy. Errors of the measurement have been analyzed. (C) 2001 Society of Photo-Optical Instrumentation Engineers.

Structural requirements for catalysis: studies on RTEM-1 β-lactamase

β-lactamases are a group of enzymes that confer resistance to penam and cephem antibiotics by hydrolysis of the β-lactam ring, thereby inactivating the antibiotic. Crystallographic and computer modeling studies of RTEM-1 β-lactamase have indicated that Asp 132, a strictly conserved residue among the class A β-lactamases, appears to be involved in substrate binding, catalysis, or both. To study the contribution of residue 132 to β-lactamase function, site saturation mutagenesis was used to generate mutants coding for all 20 amino acids at position 132. Phenotypic screening of all mutants indicated that position 132 is very sensitive to amino acid changes, with only N132C, N132D, N132E, and N132Q showing any appreciable activity. Kinetic analysis of three of these mutants showed increases in K_M, along with substantial decreases in k_(cat). Efforts to trap a stable acyl-enzyme intermediate were unsuccessfuL These results indicate that residue 132 is involved in substrate binding, as well as catalysis, and supports the involvement of this residue in acylation as suggested by Strynadka et al.

Crystallographic and computer modeling studies of RTEM-1 β-lactamase have indicated that Lys 73 and Glu 166, two strictly conserved residues among the class A β-lactamases, appear to be involved in substrate binding, catalysis, or both. To study the contribution of these residues to β-lactamase function, site saturation mutagenesis was used to generate mutants coding for all 20 amino acids at positions 73 and 166. Then all 400 possible combinations of mutants were created by combinatorial mutagenesis. The colonies harboring the mutants were screened for growth in the presence of ampicillin. The competent colonys' DNA were sequenced, and kinetic parameters investigated. It was found that lysine is essential at position 73, and that position 166 only tolerated fairly conservative changes (Aspartic acid, Histidine, and Tyrosine). These functional mutants exhibited decreased kcat's, but K_M was close to wild-type levels. The results of the combinatorial mutagenesis experiments indicate that Lysis absolutely required for activity at position 73; no mutation at residue 166 can compensate for loss of the long side chain amine. The active mutants found--K73K/E166D, K73KIE166H, and K73KIE166Y were studied by kinetic analysis. These results reaffirmed the function of residue 166 as important in catalysis, specifically deacylation.

The identity of the residue responsible for enhancing the active site serine (Ser 70) in RTEM-1 β-lactamase has been disputed for some time. Recently, analysis of a crystal structure of RTEM-1 β-lactamase with covalently bound intermediate was published, and it was suggested that Lys 73, a strictly conserved residue among the class A β-lactamases, was acting as a general base, activating Ser 70. For this to be possible, the pK_a of Lys 73 would have to be depressed significantly. In an attempt to assay the pK_a of Lys 73, the mutation K73C was made. This mutant protein can be reacted with 2-bromoethylamine, and activity is restored to near wild type levels. ^(15)N-2-bromoethylamine hydrobromide and ^(13)C-2-bromoethylamine hydrobromide were synthesized. Reacting these compounds with the K73C mutant gives stable isotopic enrichment at residue 73 in the form of aminoethylcysteine, a lysine homologue. The pK_a of an amine can be determined by NMR titration, following the change in chemical shift of either the ^(15)N-amine nuclei or adjacent Be nuclei as pH is changed. Unfortunately, low protein solubility, along with probable label scrambling in the Be experiment, did not permit direct observation of either the ^(15)N or ^(13)C signals. Indirect detection experiments were used to observe the protons bonded directly to the ^(13)C atoms. Two NMR signals were seen, and their chemical shift change with pH variation was noted. The peak which was determined to correspond to the aminoethylcysteine residue shifted from 3.2 ppm down to 2.8 ppm over a pH range of 6.6 to 12.5. The pK_a of the amine at position 73 was determined to be ~10. This indicates that residue 73 does not function as a general base in the acylation step of the reaction. However the experimental measurement takes place in the absence of substrate. Since the enzyme undergoes conformational changes upon substrate binding, the measured pK_a of the free enzyme may not correspond to the pK_a of the enzyme substrate complex.

Las residencias señoriales bajomedievales en el País Vasco: análisis de la evolución del sistema constructivo y sus elementos singulares

8 pag.

Prevalence and magnitude of body weight and image dissatisfaction among women in dietetics majors

[EN] Some authors have suggested that body weight dissatisfaction may be high in students majoring in dietetics. Therefore, this study was conducted to examine the extent of body weight and image dissatisfaction in a sample of women in dietetics major. Additionally, predictors of magnitude of body weight dissatisfaction were analyzed. Participants were 62 volunteers with normalweight whose mean age was 21.87±1.89 years old (nonrandom sample). The assessment instruments included anthropometric measurements, a somatomorphic matrix test and an eating disorders inventory (EDI-2). Data were analyzed using SPSS vs. 15.0. A larger proportion of students chose an ideal body weight lower than actual weight (67.7%) and body image with less body fat and more muscle mass than actual values (56.4%). The magnitude of body weight dissatisfaction was associated with muscle mass and body fat dissatisfaction, and with the subscale of EDI-2 “body dissatisfaction”. So, from a public health standpoint, we consider important to continue working in this line of research with the aim of better understanding the extent of body weight dissatisfaction in women dietitians, and how this dissatisfaction could interfere with their professional practice.

Completions of epsilon-dense partial Latin squares

A classical question in combinatorics is the following: given a partial Latin square $P$, when can we complete $P$ to a Latin square $L$? In this paper, we investigate the class of textbf{$epsilon$-dense partial Latin squares}: partial Latin squares in which each symbol, row, and column contains no more than $epsilon n$-many nonblank cells. Based on a conjecture of Nash-Williams, Daykin and H"aggkvist conjectured that all $frac{1}{4}$-dense partial Latin squares are completable. In this paper, we will discuss the proof methods and results used in previous attempts to resolve this conjecture, introduce a novel technique derived from a paper by Jacobson and Matthews on generating random Latin squares, and use this novel technique to study $ epsilon$-dense partial Latin squares that contain no more than $delta n^2$ filled cells in total.

In Chapter 2, we construct completions for all $ epsilon$-dense partial Latin squares containing no more than $delta n^2$ filled cells in total, given that $epsilon < frac{1}{12}, delta < frac{ left(1-12epsilonright)^{2}}{10409}$. In particular, we show that all $9.8 cdot 10^{-5}$-dense partial Latin squares are completable. In Chapter 4, we augment these results by roughly a factor of two using some probabilistic techniques. These results improve prior work by Gustavsson, which required $epsilon = delta leq 10^{-7}$, as well as Chetwynd and H"aggkvist, which required $epsilon = delta = 10^{-5}$, $n$ even and greater than $10^7$.

If we omit the probabilistic techniques noted above, we further show that such completions can always be found in polynomial time. This contrasts a result of Colbourn, which states that completing arbitrary partial Latin squares is an NP-complete task. In Chapter 3, we strengthen Colbourn's result to the claim that completing an arbitrary $left(frac{1}{2} + epsilonright)$-dense partial Latin square is NP-complete, for any $epsilon > 0$.

Colbourn's result hinges heavily on a connection between triangulations of tripartite graphs and Latin squares. Motivated by this, we use our results on Latin squares to prove that any tripartite graph $G = (V_1, V_2, V_3)$ such that begin{itemize} item $|V_1| = |V_2| = |V_3| = n$, item For every vertex $v in V_i$, $deg_+(v) = deg_-(v) geq (1- epsilon)n,$ and item $|E(G)| > (1 - delta)cdot 3n^2$ end{itemize} admits a triangulation, if $epsilon < frac{1}{132}$, $delta < frac{(1 -132epsilon)^2 }{83272}$. In particular, this holds when $epsilon = delta=1.197 cdot 10^{-5}$.

This strengthens results of Gustavsson, which requires $epsilon = delta = 10^{-7}$.

In an unrelated vein, Chapter 6 explores the class of textbf{quasirandom graphs}, a notion first introduced by Chung, Graham and Wilson cite{chung1989quasi} in 1989. Roughly speaking, a sequence of graphs is called "quasirandom"' if it has a number of properties possessed by the random graph, all of which turn out to be equivalent. In this chapter, we study possible extensions of these results to random $k$-edge colorings, and create an analogue of Chung, Graham and Wilson's result for such colorings.

Ground-state energy shifts of H-like Ti under dense and hot plasma conditions

In this study, by adopting the ion sphere model, the self-consistent. field method is used with the Poisson-Boltzmann equation and the Dirac equation to calculate the ground-state energies of H-like Ti at a plasma electron density from 10(22) cm(-3) to 10(24) cm(-3) and the electron temperature from 100 eV to 3600 eV. The ground-state energy shifts of H-like Ti show different trends with the electron density and the electron temperature. It is shown that the energy shifts increase with the increase in the electron density and decrease with the increase in the electron temperature. The energy shifts are sensitive to the electron density, but only sensitive to the low electron temperature. In addition, an accurately fitting formula is obtained to fast estimate the ground-state energies of H-like Ti. Such fitted formula can also be used to estimate the critical electron density of pressure ionization for the ground state of H-like Ti.

Ari progresiboaz: euskararen kasu markak

Pablo Albizu eta Beatriz Fernández (arg.)

Investigations of DNA-mediated protein oxidation

DNA charge transport (CT) involves the efficient transfer of electrons or electron holes through the DNA π-stack over long molecular distances of at least 100 base-pairs. Despite this shallow distance dependence, DNA CT is sensitive to mismatches or lesions that disrupt π-stacking and is critically dependent on proper electronic coupling of the donor and acceptor moieties into the base stack. Favorable DNA CT is very rapid, occurring on the picosecond timescale. Because of this speed, electron holes equilibrate along the DNA π-stack, forming a characteristic pattern of DNA damage at low oxidation potential guanine multiplets. Furthermore, DNA CT may be used in a biological context. DNA processing enzymes with 4Fe4S clusters can perform DNA-mediated electron transfer (ET) self-exchange reactions with other 4Fe4S cluster proteins, even if the proteins are quite dissimilar, as long as the DNA-bound [4Fe4S]^3+/2+ redox potentials are conserved. This mechanism would allow low copy number DNA repair proteins to find their lesions efficiently within the cell. DNA CT may also be used biologically for the long-range, selective activation of redox-active transcription factors. Within this work, we pursue other proteins that may utilize DNA CT within the cell and further elucidate aspects of the DNA-mediated ET self-exchange reaction of 4Fe4S cluster proteins.

Dps proteins, bacterial mini-ferritins that protect DNA from oxidative stress, are implicated in the survival and virulence of pathogenic bacteria. One aspect of their protection involves ferroxidase activity, whereby ferrous iron is bound and oxidized selectively by hydrogen peroxide, thereby preventing formation of damaging hydroxyl radicals via Fenton chemistry. Understanding the specific mechanism by which Dps proteins protect the bacterial genome could inform the development of new antibiotics. We investigate whether DNA-binding E. coli Dps can utilize DNA CT to protect the genome from a distance. An intercalating ruthenium photooxidant was employed to generate oxidative DNA damage via the flash-quench technique, which localizes to a low potential guanine triplet. We find that Dps loaded with ferrous iron, in contrast to Apo-Dps and ferric iron-loaded Dps which lack available reducing equivalents, significantly attenuates the yield of oxidative DNA damage at the guanine triplet. These data demonstrate that ferrous iron-loaded Dps is selectively oxidized to fill guanine radical holes, thereby restoring the integrity of the DNA. Luminescence studies indicate no direct interaction between the ruthenium photooxidant and Dps, supporting the DNA-mediated oxidation of ferrous iron-loaded Dps. Thus DNA CT may be a mechanism by which Dps efficiently protects the genome of pathogenic bacteria from a distance.

Further work focused on spectroscopic characterization of the DNA-mediated oxidation of ferrous iron-loaded Dps. X-band EPR was used to monitor the oxidation of DNA-bound Dps after DNA photooxidation via the flash-quench technique. Upon irradiation with poly(dGdC)₂, a signal arises with g = 4.3, consistent with the formation of mononuclear high-spin Fe(III) sites of low symmetry, the expected oxidation product of Dps with one iron bound at each ferroxidase site. When poly(dGdC)₂ is substituted with poly(dAdT)₂, the yield of Dps oxidation is decreased significantly, indicating that guanine radicals facilitate Dps oxidation. The more favorable oxidation of Dps by guanine radicals supports the feasibility of a long-distance protection mechanism via DNA CT where Dps is oxidized to fill guanine radical holes in the bacterial genome produced by reactive oxygen species.

We have also explored possible electron transfer intermediates in the DNA-mediated oxidation of ferrous iron-loaded Dps. Dps proteins contain a conserved tryptophan residue in close proximity to the ferroxidase site (W52 in E. coli Dps). In comparison to WT Dps, in EPR studies of the oxidation of ferrous iron-loaded Dps following DNA photooxidation, W52Y and W52A mutants were deficient in forming the characteristic EPR signal at g = 4.3, with a larger deficiency for W52A compared to W52Y. In addition to EPR, we also probed the role of W52 Dps in cells using a hydrogen peroxide survival assay. Bacteria containing W52Y Dps survived the hydrogen peroxide challenge more similarly to those containing WT Dps, whereas cells with W52A Dps died off as quickly as cells without Dps. Overall, these results suggest the possibility of W52 as a CT hopping intermediate.

DNA-modified electrodes have become an essential tool for the study of the redox chemistry of DNA processing enzymes with 4Fe4S clusters. In many cases, it is necessary to investigate different complex samples and substrates in parallel in order to elucidate this chemistry. Therefore, we optimized and characterized a multiplexed electrochemical platform with the 4Fe4S cluster base excision repair glycosylase Endonuclease III (EndoIII). Closely packed DNA films, where the protein has limited surface accessibility, produce EndoIII electrochemical signals sensitive to an intervening mismatch, indicating a DNA-mediated process. Multiplexed analysis allowed more robust characterization of the CT-deficient Y82A EndoIII mutant, as well as comparison of a new family of mutations altering the electrostatics surrounding the 4Fe4S cluster in an effort to shift the reduction potential of the cluster. While little change in the DNA-bound midpoint potential was found for this family of mutants, likely indicating the dominant effect of DNA-binding on establishing the protein redox potential, significant variations in the efficiency of DNA-mediated electron transfer were apparent. On the basis of the stability of these proteins, examined by circular dichroism, we proposed that the electron transfer pathway in EndoIII can be perturbed not only by the removal of aromatic residues but also through changes in solvation near the cluster.

While the 4Fe4S cluster of EndoIII is relatively insensitive to oxidation and reduction in solution, we have found that upon DNA binding, the reduction potential of the [4Fe4S]^3+/2+ couple shifts negatively by approximately 200 mV, bringing this couple into a physiologically relevant range. Demonstrated using electrochemistry experiments in the presence and absence of DNA, these studies do not provide direct molecular evidence for the species being observed. Sulfur K-edge X-ray absorbance spectroscopy (XAS) can be used to probe directly the covalency of iron-sulfur clusters, which is correlated to their reduction potential. We have shown that the Fe-S covalency of the 4Fe4S cluster of EndoIII increases upon DNA binding, stabilizing the oxidized [4Fe4S]³⁺ cluster, consistent with a negative shift in reduction potential. The 7% increase in Fe-S covalency corresponds to an approximately 150 mV shift, remarkably similar to DNA electrochemistry results. Therefore we have obtained direct molecular evidence for the shift in 4Fe4S reduction potential of EndoIII upon DNA binding, supporting the feasibility of our model whereby these proteins can utilize DNA CT to cooperate in order to efficiently find DNA lesions inside cells.

In conclusion, in this work we have explored the biological applications of DNA CT. We discovered that the DNA-binding bacterial ferritin Dps can protect the bacterial genome from a distance via DNA CT, perhaps contributing to pathogen survival and virulence. Furthermore, we optimized a multiplexed electrochemical platform for the study of the redox chemistry of DNA-bound 4Fe4S cluster proteins. Finally, we have used sulfur K-edge XAS to obtain direct molecular evidence for the negative shift in 4Fe4S cluster reduction potential of EndoIII upon DNA binding. These studies contribute to the understanding of DNA-mediated protein oxidation within cells.

Estratégia para validação de processos de esterilização por vapor saturado em autoclaves nas indústrias farmacêuticas

O objetivo deste trabalho foi criar uma metodologia de validação e revalidação dos processos de esterilização por calor úmido em autoclaves horizontais, destacando os pontos críticos do processo e concentrando esforços onde são realmente necessários. Foram realizados estudos de distribuição térmica, de penetração de calor e de desafio microbiológico na validação da autoclave STERIS FINNAQUA 6912. Com o objetivo de avaliar o impacto de uma mudança e compreender a relação entre os fatores e suas interações para o processo de esterilização, foi utilizado o planejamento fatorial 23 dos fatores densidade da carga (quantidade de itens), embalagem do produto e localização na câmara interna. Os estudos de distribuição térmica confirmaram a distribuição homogênea de calor na câmara interna durante o tempo de exposição a 121C. As temperaturas variaram entre 120,35C e 120,92C com desvio padrão máximo de 0,12C. Os estudos de penetração de calor confirmaram exposições equivalentes a 121C por 24 minutos em todos os itens da carga (F0 > 24 minutos). Em todos os estudos para cargas secas, os índices de capacidade do processo (Cpk) foram maiores que 1,33. Os ensaios de desafio microbiológico garantiram níveis de esterilidade (S.A.L.) maiores que 12 reduções logarítmicas em relação aos indicadores biológicos Geobacillus stearothermophilus. Não foi detectada a presença de endosporos sobreviventes nos 132 indicadores biológicos utilizados nos quatro ciclos desafiados. Com base no planejamento experimental verificou-se que, para o nível de significância de 95% , as mudanças nos fatores posição, embalagem e quantidade da carga não são significativas para o processo de esterilização, em autoclave com remoção forçada de ar. Já para o nível de significância de 90%, a interação Posição x Embalagem apresentou significância estatística no processo de esterilização com valor P de 0,080