Part I: The abortive infection of bacteriophage øx174 at low temperatures. Part II: The early stages in the process of infection by bacteriophage øx174

Part I

The infection of E. coli by ΦX174 at 15°C is abortive; the cells are killed by the infection but neither mature phage nor SS (single-stranded) DNA are synthesized. Parental RF (replicative form) is formed and subsequently replicated at 15°C. The RF made at 15°C shows normal infectivity and full competence to act as precursor to progeny SS DNA after an increase in temperature to 37°C. The investigations suggest that all of the proteins required for SS DNA synthesis and phage maturation are present in the abortive infection at 15°C.

Three possible causes are suggested for the abortive infection at 15°C: (a) A virus-coded protein whose role is essential to the infection is made at 15°C and assumes its native conformation, but its rate of activity is too low at this temperature to sustain the infection process. (b) Virus maturation may involve the formation of a DNA-protein complex and conformational changes which have an energy threshold infrequently reached at 15°C. (c) A host-coded protein present in uninfected cells, and whose activity is essential to the infection at all temperatures, but not to the host at 15°C, is inactive at 15°C. An hypothesis of this type is offered which proposes that the temperature-limiting factor in SS DNA synthesis in vivo may reflect a temperature-dependent property of the host DNA polymerase.

Part II

Three distinct stages are demonstrated in the process whereby ΦX174 invades its host: (1) Attachment: The phage attach to the cell in a manner that does not irreversibly alter the phage particle and which exhibits "single-hit" kinetics. The total charge on the phage particle is demonstrated to be important in determining the rate at which stable attachment is effected. The proteins specified by ΦX cistrons II, III and VII play roles, which may be indirect, in the attachment reaction. (2) Eclipse: 'The attached phage undergo a conformational change. Some of the altered phage particles spontaneously detach from the cell (in a non-infective form) while the remainder are more tightly bound to the cell. The altered phage particles detached (spontaneously or chemically) from such complexes have at least 40% of their DNA extruded from the phage coat. It is proposed that this particle is, or derives from, a direct intermediate in the penetration of the viral DNA.

The kinetics for the eclipse of attached phage particles are first-order with respect to phage concentration and biphasic; about 85% of the phage eclipse at one rate (k = 0.86 min^-1) and the remainder do so at a distinctly lesser rate (k = 0.21 min^-1).

The eclipse event is very temperature-dependent and has the relatively high Arrhenius activation energy of 36.6 kcal/mole, indicating the cooperative nature of the process. The temperature threshold for eclipse is 17 to 18°C.

At present no specific ΦX cistron is identified as affecting the eclipse process. (3) DNA penetration: A fraction of the attached, eclipsed phage particles corresponding in number to the plaque-forming units complete DNA penetration. The penetrated DNA is found in the cell as RF, and the empty phage protein coat remains firmly attached to the exterior of the cell. This step is inhibited by prior irradiation of the phage with relatively high doses of UV light and is insensitive to the presence of KCN and NaN₃. Temporally excluded superinfecting phages do not achieve DNA penetration.

Both eclipsed phage particles and empty phage protein coats may be dissociated from infected cells; some of their properties are described.

Simulation of growth process of thin film on non-planar substrate

In (2 + 1) dimension, growth process of thin film on non-planar substrate in Kuramoto-Sivashinsky model is studied with numerical simulation approach. 15 x 15 semi-ellipsoids arranged orderly on the surface of substrate are used to represent initial rough surface. The results show that at the initial stage of growth process, the surface morphology of thin film appears to be grid-structure, and the interface width constantly decreases with the growth time, then reaches minimum. However, the grid-structure becomes ambiguous, and granules of different sizes distribute evenly on the surface of thin film with the increase of growth time. Thereafter, the average size of granules and the interface width gradually increase, and the surface morphology of thin film presents fractal properties. The numerical results of height-height correlation functions of thin film verify the surface morphology of thin film to be fractal for a longer growth time. By fitting of the height-height correlation functions of thin film with different growth times, the growth process is described quantitatively. (c) 2004 Elsevier B.V. All rights reserved.

Caracterização da expressão gênica dos receptores colinérgicos nicotínicos (CHRNs) no epitélio esofágico normal e em carcinoma epidermóide de esôfago

O câncer de esôfago (CE) é uma doença extremamente agressiva e é um dos tumores mais incidentes e letais no Brasil e no mundo, sendo o carcinoma epidermóide de esôfago (CEE) o principal subtipo histológico, apresentando como principais fatores de risco o etilismo e o tabagismo na população ocidental. A exposição concomitante desses dois fatores representa um risco multiplicador para o desenvolvimento de CEE, sendo que o fumo parece ter um papel importante tanto na iniciação quanto na promoção do tumor, enquanto o álcool teria um papel mais relevante na promoção. Componentes do tabaco, como a nicotina e as nitrosaminas são potentes carcinógenos e agonistas de alta afinidade dos receptores colinérgicos nicotínicos (CHRNs), podendo atuar ativando vias de sinalização celular fundamentais para a progressão tumoral. Pouco se sabe sobre a expressão e regulação dos CHRNs na mucosa esofágica e no processo de carcinogênese desse tecido. Assim, o objetivo desse trabalho foi analisar a expressão gênica dos CHRNs no epitélio esofágico normal e em CEE, bem como sua regulação pelos fatores de risco associados ao tumor. Foi observado que as subunidades α3, α5, α7 e β4 são expressas no epitélio esofágico saudável humano enquanto as subunidades α1, α4, α9 e α10 apresentaram baixa ou nenhuma expressão nesse mesmo tecido. Além disso, foram encontradas diferenças de expressão das subunidades α3 e α7 em indivíduos etilistas e tabagistas quando comparados com indivíduos não-etilistas (subunidade α3) e não-tabagistas (subunidade α7). Nas amostras de CEE, as subunidades CHRNA5 e CHRNA7 foram encontradas superexpressas no tumor quando comparado ao tecido normal adjacente e observou-se diferença de expressão da subunidade α7 no tumor comparado com o tecido saudável e a subunidade β4 apresentou-se mais expressa no tecido tumoral e no tecido normal adjacente ao tumor do que no epitélio esofágico saudável. Entretanto, não foram encontradas diferenças de expressão de nenhuma das subunidades avaliadas nas linhagens CEE quando submetidas a tratamento com nicotina ou etanol. Os resultados obtidos sugerem uma participação dos CHRNs na fisiologia do epitélio esofágico e que os fatores de risco associados ao desenvolvimento de CEE parecem ser capazes de afetar a expressão desses receptores no epitélio esofágico.

An Ultra Low Power Non-volatile Memory in Standard CMOS Process for Passive RFID Tags

An ultra low power non-volatile memory is designed in a standard CMOS process for passive RFID tags. The memory can operate in a new low power operating scheme under a wide supply voltage and clock frequency range. In the charge pump circuit the threshold voltage effect of the switch transistor is almost eliminated and the pumping efficiency of the circuit is improved. An ultra low power 192-bit memory with a register array is implemented in a 0.18 mu M standard CMOS process. The measured results indicate that, for the supply voltage of 1.2 volts and the clock frequency of 780KHz, the current consumption of the memory is 1.8 mu A (3.6 mu A) at the read (write) rate of 1.3Mb/s (0.8Kb/s).

Ignition of single coal particle in a hot furnace under normal- and micro-gravity condition

An experimental study on ignition and combustion of single particles was conducted at normal gravity (1-g) and microgravity (l-g) for three high volatile coals with initial diameter of 1.5 and 2.0 mm, respectively. The non-intrusive twin-color pyrometry method was used to retrieve the surface temperature of the coal particle through processing the images taken by a color CCD camera. At the same time, a mathematical model considering thermal conduction inside the coal particle was developed to simulate the ignition process. Both experiments and modeling found that ignition occurred homogeneously at the beginning and then heterogeneously for the testing coal particles burning at l-g. Experimental results confirmed that ignition temperature decreased with increasing volatile content and increasing particle size. However, contradicted to previous studies, this study found that for a given coal with certain particle size, ignition temperature was about 50–80 K lower at l-g than that at 1-g. The model predictions agreed well with the l-g experimental data on ignition temperature. The criterion that the temperature gradient in the space away from the particle surface equaled to zero was validated to determine the commence of homogeneous ignition. Thermal conduction inside the particle could have a noticeable effect for determining the ignition temperature. With the consideration of thermal conduction, the critical size for the phase transient from homogeneous to heterogeneous is about 700 lm at ambient temperature 1500 K and oxygen concentration 0.23. 2009 The Combustion Institute. Published by Elsevier Inc. All rights reserved.

Non-product related environmental process and production methods (NPR-PPMs): a view from international trade law and international environmental governance

The primary aim of this thesis is to analyse legal and governance issues in the use of Environmental NPR-PPMs, particularly those aiming to promote sustainable practices or to protect natural resources. NPR-PPMs have traditionally been thought of as being incompatible with the rules of the World Trade Organization (WTO). However, the issue remains untouched by WTO adjudicatory bodies. One can suggest that WTO adjudicatory bodies may want to leave this issue to the Members, but the analysis of the case law also seems to indicate that the question of legality of NPR-PPMs has not been brought ‘as such’ in dispute settlement. This thesis advances the argument that despite the fact that the legal status of NPR-PPMs remains unsettled, during the last decades adjudicatory bodies have been scrutinising environmental measures based on NPR-PPMs just as another expression of the regulatory autonomy of the Members. Though NPR-PPMs are regulatory choices associated with a wide range of environmental concerns, trade disputes giving rise to questions related to the legality of process-based measures have been mainly associated with the protection of marine wildlife (i.e., fishing techniques threatening or affecting animal species). This thesis argues that environmental objectives articulated as NPR-PPMs can indeed qualify as legitimate objectives both under the GATT and the TBT Agreement. However, an important challenge for the their compatibility with WTO law relate to aspects associated with arbitrary or unjustifiable discrimination. In the assessment of discrimination procedural issues play an important role. This thesis also elucidates other important dimensions to the issue from the perspective of global governance. One of the arguments advanced in this thesis is that a comprehensive analysis of environmental NPR-PPMs should consider not only their role in what is regarded as trade barriers (governmental and market-driven), but also their significance in global objectives such as the transition towards a green economy and sustainable patterns of consumption and production.

A non-Newtonian computational fluid dynamics study of the stencil printing process

This paper describes the application of computational fluid dynamics (CFD) to simulate the macroscopic bulk motion of solder paste ahead of a moving squeegee blade in the stencil printing process during the manufacture of electronic components. The successful outcome of the stencil printing process is dependent on the interaction of numerous process parameters. A better understanding of these parameters is required to determine their relation to print quality and improve guidelines for process optimization. Various modelling techniques have arisen to analyse the flow behaviour of solder paste, including macroscopic studies of the whole mass of paste as well as microstructural analyses of the motion of individual solder particles suspended in the carrier fluid. This work builds on the knowledge gained to date from earlier analytical models and CFD investigations by considering the important non-Newtonian rheological properties of solder pastes which have been neglected in previous macroscopic studies. Pressure and velocity distributions are obtained from both Newtonian and non-Newtonian CFD simulations and evaluated against each other as well as existing established analytical models. Significant differences between the results are observed, which demonstrate the importance of modelling non-Newtonian properties for realistic representation of the flow behaviour of solder paste.

A novel shrinkage technique based on the normal inverse Gaussian density model

This paper proposes a novel image denoising technique based on the normal inverse Gaussian (NIG) density model using an extended non-negative sparse coding (NNSC) algorithm proposed by us. This algorithm can converge to feature basis vectors, which behave in the locality and orientation in spatial and frequency domain. Here, we demonstrate that the NIG density provides a very good fitness to the non-negative sparse data. In the denoising process, by exploiting a NIG-based maximum a posteriori estimator (MAP) of an image corrupted by additive Gaussian noise, the noise can be reduced successfully. This shrinkage technique, also referred to as the NNSC shrinkage technique, is self-adaptive to the statistical properties of image data. This denoising method is evaluated by values of the normalized signal to noise rate (SNR). Experimental results show that the NNSC shrinkage approach is indeed efficient and effective in denoising. Otherwise, we also compare the effectiveness of the NNSC shrinkage method with methods of standard sparse coding shrinkage, wavelet-based shrinkage and the Wiener filter. The simulation results show that our method outperforms the three kinds of denoising approaches mentioned above.

Neurokinin A is the predominant tachykinin in human bronchoalveolar lavage fluid in normal and asthmatic subjects

BACKGROUND—Multiple sensory neuropeptides are present in human airways and may contribute to diseases such as asthma. This study quantified and characterised substance P (SP), neurokinin A (NKA), and calcitonin gene related peptide (CGRP) immunoreactivity in bronchoalveolar lavage fluid in asthmatic and normal subjects.
METHODS—Using specific radioimmunoassay (RIA), SP, NKA and CGRP were measured in bronchoalveolar lavage fluid from asthmatic subjects (n = 5), normal subjects (n = 5), atopic non-asthmatic subjects (n = 6), and asthmatic subjects four hours after allergen challenge (n = 12). Peptide immunoreactivity was characterised using high performance liquid chromatography (HPLC) and RIA.
RESULTS—No SP or CGRP immunoreactivity was detected in any of the fractions from samples after extraction, HPLC, and RIA. Non-specific binding resulted in spurious SP immunoreactivity being detected in bronchoalveolar lavage fluid when no extraction process was employed. NKA was detected in significant amounts in asthmatic (median 550, range 425-625 pg/ml) and normal subjects (median 725, range 350-1425 pg/ml). The level of NKA was significantly higher in the asthmatic subjects after allergen challenge (median 750, range 350-1250 pg/ml) than in unchallenged asthmatic subjects (median 600, range 425-600 pg/ml, p<0.01).
CONCLUSIONS—Extraction and characterisation of peptides from bronchoalveolar lavage fluid must be performed to ensure that the measured immunoreactivity represents target peptide. NKA is present in bronchoalveolar lavage fluid in high concentrations and is the predominant tachykinin. The concentrations of NKA are similar in normal subjects and subjects with mild asthma.

Investigating paediatric airways by non-bronchoscopic lavage: Normal cellular data

Background: Bronchoscopic bronchoalveolar lavage in children to investigate bronchia disorders such as asthtna has both ethical and procedural difficulties.


Objective: The aim of this study was to establish a standardized non-bronchoscopic method to perform bronchoalveolar lavage in children attending for elective surgery to obtain normal cellular data.


Methods: Bronchoalveolar lavage was performed on normal children (n= 55) by infusing saline (20 mL) through an 8 FG suction catheter passed after endotracheal intubation. Oxygen saturation, heart and respiratory rate were monitored during the bronchoalveolar lavage procedure. Cellular analysis and total protein estimation of the lavage fluid were performed. Epithelial lining fluid volume was calculated (n = 15) using the urea dilution method.


Results: The procedure was well tolerated by all children. Total cell count and differential cell count for children (macrophages 70.8 ± 2.3%, lymphocytes 3.8 ± 0.6%, neutrophils 5,7 ± 1.0%, eosinophils 0.14 ± 0.03%. epithelial cells 19.6 ± 2.1%, mast cells 0.21 ± 0.02%) were similar to those reported for adults. Age and sex comparisons revealed no differences between groups. The mean total protein recovered in the cell free supernatant was 49.72 ± 4.29 mg/L and epithelial lining fluid volume was 0.82 ± 0.11% of return lavageate.


Conclusion This method allows bronchoalveolar lavage to be performed safely and quickly on children attending for routine elective surgery. Using this method and taking the ‘window of opportunity’ of elective surgery, the presence or absence of airway inflammation could be studied in children with various patterns of asthma during relatively asymptomatic periods.