Limitations of HLA-transgenic mice in presentation of HLA-restricted cytotoxic T-cell epitopes from endogenously processed human papillomavirus type 16 E7 protein

We investigated the use of mice transgenic for human leucocyte antigen (HLA) A*0201 antigen-binding domains to test vaccines composed of defined HLA A*0201-restricted cytotoxic T-lymphocyte (CTL) epitopes of human papillomavirus (HPV) type 16 E7 oncoprotein. HPV is detected in >90% of cervical carcinomas. HPV16 E7 oncoprotein transforms cells of the uterine cervix and functions as a tumour-associated antigen to which immunotherapeutic strategies may be directed. We report that although the HLA A*0201 E7 epitope peptides function both to prime for E7 CTL responses, and to sensitize target cells for E7-directed CTL killing in situations where antigen processing is not required, the epitopes are not processed out of either endogenously expressed or immunization-introduced E7, by the mouse antigen-processing and presentation machinery. Thus (1) CTL induced by HLA A*0201 peptide immunization killed E7 peptide-pulsed target cells, but did not kill target cells expressing whole E7; (2) immunization with whole E7 protein did not elicit CTL directed to HLA A*0201-restricted E7 CTL epitopes; (3) HLA A*0201-restricted CTL epitopes expressed in the context of a DNA polytope vaccine did not activate E7-specific T cells either in 'conventional' HLA A*0201 transgenic (A2.1K(b) ) mice, or in HHD transgenic mice in which expression of endogenous H-2 class 1 is precluded; and (4) HLA A*0201 E7 peptide epitope immunization was incapable of preventing the growth of an HLA A*0201- and E7-expressing tumour. There are generic implications for the universal applicability of HLA-class 1 transgenic mice for studies of human CTL epitope presentation in murine models of human infectious disease where recognition of endogenously processed antigen is necessary. There are also specific implications for the use of HLA A2 transgenic mice for the development of E7-based therapeutic vaccines for cervical cancer.

Influence of planting date, method of handling and seed size on supersweet sweet corn seed quality

A supersweet sweet corn hybrid, Pacific H5, was planted at weekly intervals (P-1 to P-5) in spring in South-Eastern Queensland. All plantings were harvested at the same time resulting in immature seed for the last planting (P-5). The seed was handled by three methods: manual harvest and processing (M-1), manual harvest and mechanical processing (M-2) and mechanical harvest and processing (M-3), and later graded into three sizes (small, medium and large). After eight months storage at 12-14degreesC, seed was maintained at 30degreesC with bimonthly monitoring of germination for fourteen months and seed damage at the end of this period. Seed quality was greatest for M-1 and was reduced by mechanical processing but not by mechanical harvesting. Large and medium seed had higher germination due to greater storage reserves but also more seed damage during mechanical processing. Immature seed from premature harvest (P-5) had poor quality especially when processed mechanically and reinforced the need for harvested seed to be physiologically mature.

Four cytotoxic N4-substituted thiosemicarbazones derived from 2-hydroxynaphthalene-1-carboxaldehyde

The X-ray crystal structures are reported of four novel and potentially O,N,S-tridentate donor ligands that demonstrate antitumour activity. These ligands are 1-[(4-methylthiosemicarbazono)methyl]-2-naphthol, C13H13N3OS, (III), 1-[(4-ethylthiosemicarbazono)methyl]-2-naphthol, C14H15N3OS, (IV), 1-[(4-phenylthiosemicarbazono)methyl]-2-naphthol, C18H15N3OS, (V), and 1-[(4,4-dimethylthiosemicarbazono)methyl]-2-naphthol dimethyl sulfoxide solvate, C14H15N3OS.C2H6OS, (VI). These chelators are N4-substituted thiosemicarbazones, each based on the same parent aldehyde, namely 2-zhydroxynaphthalene-1-carboxaldehyde isonicotinoylhydrazone. Conformational variations within this series are discussed in relation to the optimum conformation for metal-ion binding.

Cytotoxic iron chelators: Characterization of the structure, solution chemistry and redox activity of ligands and iron complexes of the di-2-pyridyl ketone isonicotinoyl hydrazone (HPKIH) analogues

Di-2-pyridyl ketone isonicotinoyl hydrazone (HPKIH) and a range of its analogues comprise a series of monobasic acids that are capable of binding iron (Fe) as tridentate (N,N,O) ligands. Recently, we have shown that these chelators are highly cytotoxic, but show selective activity against cancer cells. Particularly interesting was the fact that cytotoxicity of the HPKIH analogues is maintained even after complexation with Fe. To understand the potent anti-tumor activity of these compounds, we have fully characterized their chemical properties. This included examination of the solution chemistry and X-ray crystal structures of both the ligands and Fe complexes from this class and the ability of these complexes to mediate redox reactions. Potentiometric titrations demonstrated that all chelators are present predominantly in their charge-neutral form at physiological pH (7.4), allowing access across biological membranes. Keto-enol tautomerism of the ligands was identified, with the tautomers exhibiting distinctly different protonation constants. Interestingly, the chelators form low-spin (diamagnetic) divalent Fe complexes in solution. The chelators form distorted octahedral complexes with Fe-II, with two tridentate ligands arranged in a meridional fashion. Electrochemistry of the Fe complexes in both aqueous and non-aqueous solutions revealed that the complexes are oxidized to their ferric form at relatively high potentials, but this oxidation is coupled to a rapid reaction with water to form a hydrated (carbinolamine) derivative, leading to irreversible electrochemistry. The Fe complexes of the HPKIH analogues caused marked DNA degradation in the presence of hydrogen peroxide. This observation confirms that Fe complexes from the HPKIH series mediate Fenton chemistry and do not repel DNA. Collectively, studies on the solution chemistry and structure of these HPKIH analogues indicate that they can bind cellular Fe and enhance its redox activity, resulting in oxidative damage to vital biomolecules.

Exposure to microbial volatile organic compounds in a waste-handling unit

The production of MVOC by fungi has been taken into account especially from the viewpoint of indoor pollution with microorganisms but the relevance of fungal metabolites in working environments has not been sufficiently studied. The purpose of this study was to assess exposure to MVOCs in a waste-handling unit. It was used Multirae equipment (RAE Systems) to measured MVOCs concentration with a 10.6 eV lamps. The measurements were done near workers nose and during the normal activities. All measurements were done continuously and had the duration of 5 minutes at least. It was consider the higher value obtained in each measurement. In addition, for knowing fungi contamination, five air samples of 50 litres were collected through impaction method at 140 L/minute, at one meter tall, on to malt extract agar with the antibiotic chloramphenicol (MEA). MVOCs results range between 4.7 ppm and 8.9 ppm in the 6 locations consider. These results are eight times higher than normally obtained in indoor settings. Considering fungi results, two species were identified in air, being the genera Penicillium found in all the samples in uncountable colonies and Rhizopus only in one sample (40 UFC/m3). These fungi are known as MVOCs producers, namely terpenoids, ketones, alcohols and others. Until now, there has been no evidence that MVOCs are toxicologically relevant, but further epidemiological research is necessary to elucidate their role on human’s health, particularly in occupational settings where microbiological contamination is common. Additionally, further research should concentrate on quantitative analyses of specific MVOCs.

Evaluation of the improvements on pharmacy technicians' work practices after implementation of operational procedures

Several antineoplasic drugs have been demonstrated to be carcinogenic or to have mutagenic and teratogenic effects. The greatest protection is achieved with the implementation of administrative and engineering controls and safety procedures. Objective: to evaluate the improvements on pharmacy technicians' work practices, after the implementation of operational procedures related to individual protection, biologic safety cabinet disinfection and cytotoxic drug preparation. Method: case-study in a hospital pharmacy undergoing a certification process. Six pharmacy technicians were observed during their daily activities. Characterization of the work practices was made using a checklist based on ISOPP and PIC guidelines. The variables studied concerning cleaning/disinfection procedures, personal protective equipment and procedures for preparing cytotoxic drugs. The same work practices were evaluated after four months of operational procedures implementation. Concordance between work practices and guidelines was considered to be a quality indicator (guidelines concordance practices number/total number of practices x 100). Results: improvements were observed after operational procedures implementation. An improvement of 6,25% in personal protective equipment practice was achieved by changing second pair of gloves every thirty minutes. The major progress, 10%, was obtained in disinfection procedure, where 80% of tasks are now realized according to guidelines.By now, we hot an improvement of only 1% at drug preparation procedure by placing one cytotoxic drug at a time inside the biological safety cabinet. Then, 85% of practices are according to guidelines. Conclusion: before operational procedures implementation 80,3% of practices were according to the guidelines, while now is 84,4%. This indicates that is necessary to review the procedures frequently in the benefit to reduce the risks associated with handling cytotoxic drugs and maintenance of drug specifications.

Antimicrobial and cytotoxic assessment of marine cyanobacteria - synechocystis and synechococcus

Aqueous extracts and organic solvent extracts of isolated marine cyanobacteria strains were tested for antimicrobial activity against a fungus, Gram-positive and Gram-negative bacteria and for cytotoxic activity against primary rat hepatocytes and HL-60 cells. Antimicrobial activity was based on the agar diffusion assay. Cytotoxic activity was measured by apoptotic cell death scored by cell surface evaluation and nuclear morphology. A high percentage of apoptotic cells were observed for HL-60 cells when treated with cyanobacterial organic extracts. Slight apoptotic effects were observed in primary rat hepatocytes when exposed to aqueous cyanobacterial extracts. Nine cyanobacteria strains were found to have antibiotic activity against two Gram-positive bacteria, Clavibacter michiganensis subsp. insidiosum and Cellulomonas uda. No inhibitory effects were found against the fungus Candida albicans and Gram-negative bacteria. Marine Synechocystis and Synechococcus extracts induce apoptosis in eukaryotic cells and cause inhibition of Gram-positive bacteria. The different activity in different extracts suggests different compounds with different polarities.

A new natural spiro heterocyclic compound and the cytotoxic activity of the secondary metabolites from Juniperus brevifolia leaves

Copyright © 2010 Elsevier B.V. All rights reserved.

Code offloading on real-time multimedia systems: a framework for handling code mobility and code offloading in a QoS aware environment

Actualmente, os smartphones e outros dispositivos móveis têm vindo a ser dotados com cada vez maior poder computacional, sendo capazes de executar um vasto conjunto de aplicações desde simples programas de para tirar notas até sofisticados programas de navegação. Porém, mesmo com a evolução do seu hardware, os actuais dispositivos móveis ainda não possuem as mesmas capacidades que os computadores de mesa ou portáteis. Uma possível solução para este problema é distribuir a aplicação, executando partes dela no dispositivo local e o resto em outros dispositivos ligados à rede. Adicionalmente, alguns tipos de aplicações como aplicações multimédia, jogos electrónicos ou aplicações de ambiente imersivos possuem requisitos em termos de Qualidade de Serviço, particularmente de tempo real. Ao longo desta tese é proposto um sistema de execução de código remota para sistemas distribuídos com restrições de tempo-real. A arquitectura proposta adapta-se a sistemas que necessitem de executar periodicamente e em paralelo mesmo conjunto de funções com garantias de tempo real, mesmo desconhecendo os tempos de execução das referidas funções. A plataforma proposta foi desenvolvida para sistemas móveis capazes de executar o Sistema Operativo Android.

Assessment of genotoxic effects in nurses handling cytostatic drugs

Several antineoplastic drugs have been classified as carcinogens by the International Agency for Research on Cancer (IARC) on the basis of epidemiological findings, animal carcinogenicity data, and outcomes of in vitro genotoxicity studies. 5-Fluorouracil (5-FU), which is easily absorbed through the skin, is the most frequently used antineoplastic agent in Portuguese hospitals and therefore may be used as an indicator of surface contamination. The aims of the present investigation were to (1) examine surface contamination by 5-FU and (2) assess the genotoxic risk using cytokinesis-block micronucleus assay in nurses from two Portuguese hospitals. The study consisted of 2 groups: 27 nurses occupationally exposed to cytostatic agents (cases) and 111 unexposed individuals (controls). Peripheral blood lymphocytes (PBL) were collected in order to measure micronuclei (MN) in both groups. Hospital B showed a higher numerical level of contamination but not significantly different from Hospital A. However; Hospital A presented the highest value of contamination and also a higher proportion of contaminated samples. The mean frequency of MN was significantly higher in exposed workers compared with controls. No significant differences were found among MN levels between the two hospitals. The analysis of confounding factors showed that age is a significant variable in MN frequency occurrence. Data suggest that there is a potential genotoxic damage related to occupational exposure to cytostatic drugs in oncology nurses.

Handling mobility on a QoS-Aware servicebased framework for mobile systems

Mobile applications are becoming increasingly more complex and making heavier demands on local system resources. Moreover, mobile systems are nowadays more open, allowing users to add more and more applications, including third-party developed ones. In this perspective, it is increasingly expected that users will want to execute in their devices applications which supersede currently available resources. It is therefore important to provide frameworks which allow applications to benefit from resources available on other nodes, capable of migrating some or all of its services to other nodes, depending on the user needs. These requirements are even more stringent when users want to execute Quality of Service (QoS) aware applications, such as voice or video. The required resources to guarantee the QoS levels demanded by an application can vary with time, and consequently, applications should be able to reconfigure themselves. This paper proposes a QoS-aware service-based framework able to support distributed, migration-capable, QoS-enabled applications on top of the Android Operating system.

Handling shared resources and precedence constraints in open systems

There is an increasing demand for highly dynamic realtime systems where several independently developed applications with different timing requirements can coexist. This paper proposes a protocol to integrate shared resources and precedence constraints among tasks in such systems assuming no precise information on critical sections and computation times is available. The concept of bandwidth inheritance is combined with a capacity sharing and stealing mechanism to efficiently exchange bandwidth among needed tasks, minimising the cost of blocking.

Handling QoS dependencies in distributed cooperative real-time systems

Due to the growing complexity and adaptability requirements of real-time embedded systems, which often exhibit unrestricted inter-dependencies among supported services and user-imposed quality constraints, it is increasingly difficult to optimise the level of service of a dynamic task set within an useful and bounded time. This is even more difficult when intending to benefit from the full potential of an open distributed cooperating environment, where service characteristics are not known beforehand. This paper proposes an iterative refinement approach for a service’s QoS configuration taking into account services’ inter-dependencies and quality constraints, and trading off the achieved solution’s quality for the cost of computation. Extensive simulations demonstrate that the proposed anytime algorithm is able to quickly find a good initial solution and effectively optimises the rate at which the quality of the current solution improves as the algorithm is given more time to run. The added benefits of the proposed approach clearly surpass its reducedoverhead.

Handling random errors and biases in methods used for short-term dietary assessment

Epidemiological studies have shown the effect of diet on the incidence of chronic diseases; however, proper planning, designing, and statistical modeling are necessary to obtain precise and accurate food consumption data. Evaluation methods used for short-term assessment of food consumption of a population, such as tracking of food intake over 24h or food diaries, can be affected by random errors or biases inherent to the method. Statistical modeling is used to handle random errors, whereas proper designing and sampling are essential for controlling biases. The present study aimed to analyze potential biases and random errors and determine how they affect the results. We also aimed to identify ways to prevent them and/or to use statistical approaches in epidemiological studies involving dietary assessments.