ProxiMAX randomisation:a new technology for non-degenerate saturation mutagenesis of contiguous codons

Back in 2003, we published ‘MAX’ randomisation, a process of non-degenerate saturation mutagenesis using exactly 20 codons (one for each amino acid) or else any required subset of those 20 codons. ‘MAX’ randomisation saturates codons located in isolated positions within a protein, as might be required in enzyme engineering, or else on one face of an alpha-helix, as in zinc finger engineering. Since that time, we have been asked for an equivalent process that can saturate multiple, contiguous codons in a non-degenerate manner. We have now developed ‘ProxiMAX’ randomisation, which does just that: generating DNA cassettes for saturation mutagenesis without degeneracy or bias. Offering an alternative to trinucleotide phosphoramidite chemistry, ProxiMAX randomisation uses nothing more sophisticated than unmodified oligonucleotides and standard molecular biology reagents. Thus it requires no specialised chemistry, reagents nor equipment and simply relies on a process of saturation cycling comprising ligation, amplification and digestion for each cycle. The process can encode both unbiased representation of selected amino acids or else encode them in pre-defined ratios. Each saturated position can be defined independently of the others. We demonstrate accurate saturation of up to 11 contiguous codons. As such, ProxiMAX randomisation is particularly relevant to antibody engineering.

ProxiMAX randomisation:a new technology for non-degenerate saturation mutagenesis of contiguous codons

Back in 2003, we published ‘MAX’ randomisation, a process of non-degenerate saturation mutagenesis using exactly 20 codons (one for each amino acid) or else any required subset of those 20 codons. ‘MAX’ randomisation saturates codons located in isolated positions within a protein, as might be required in enzyme engineering, or else on one face of an alpha-helix, as in zinc finger engineering. Since that time, we have been asked for an equivalent process that can saturate multiple, contiguous codons in a non-degenerate manner. We have now developed ‘ProxiMAX’ randomisation, which does just that: generating DNA cassettes for saturation mutagenesis without degeneracy or bias. Offering an alternative to trinucleotide phosphoramidite chemistry, ProxiMAX randomisation uses nothing more sophisticated than unmodified oligonucleotides and standard molecular biology reagents. Thus it requires no specialised chemistry, reagents nor equipment and simply relies on a process of saturation cycling comprising ligation, amplification and digestion for each cycle. The process can encode both unbiased representation of selected amino acids or else encode them in pre-defined ratios. Each saturated position can be defined independently of the others. We demonstrate accurate saturation of up to 11 contiguous codons. As such, ProxiMAX randomisation is particularly relevant to antibody engineering.

Application of Network Technologies for Development of Medical Data-advisory Clinic “Med-Health”

In this article the medical data-advisory web-resource developed by authors is considered. This resource allows carrying out information interchange between consumers of medical services and the medical establishments which give these services, and firms-manufacturers of medical equipment and medicaments. Main sections of this web-site, their purposes and capabilities are considered in this article.

Properties of ultra fast deposited diamond-like hydrogenated carbon films

Hydrogenated amorphous carbon films with diamond like structures have been formed on different substrates at very low energies and temperatures by a plasma enhanced chemical vapor deposition process employing acetylene as the precursor gas. The plasma source was of a cascaded arc type with Ar as carrier gas. The films were grown at very high deposition rates. Deposition on Si, glass and plastic substrates has been studied and the films characterized in terms of sp3 content, roughness, hardness, adhesion and optical properties. Deposition rates up to 20 nm/s have been achieved at substrate temperatures below 100°C. The typical sp3 content of 60-75% in the films was determined by X-ray generated Auger electron spectroscopy. Hardness, reduced modulus and adhesion were measured using a MicroMaterials Nano Test Indenter/Scratch tester. Hardness was found to vary from 4 to 13 GPa depending on deposition conditions. Adhesion was significantly influenced by the substrate temperature and in situ DC cleaning. Hydrogen content in the film was measured by a combination of the Fourier transform infrared and Rutherford backscattering techniques. Advantages of these films are: low ion energy and deposition temperature, very high deposition rates, low capital cost of the equipment and the possibility of film properties being tailored according to the desired application.

Market definition in industrial high-technology markets

This dissertation analyzes how marketers define markets in technology-based industries. One of the most important strategic decisions marketers face is determining the optimal market for their products. Market definition is critical in dynamic high technology markets characterized by high levels of market and technological uncertainty. Building on literature from marketing and related disciplines, this research is the first in-depth study of market definition in industrial markets. Using a national, probability sample stratified by firm size, 1,000 marketing executives in nine industries (automation, biotechnology, computers, medical equipment and instrumentation, pharmaceuticals, photonics, software, subassemblies and components, and telecommunications) were surveyed via a mail questionnaire. A 20.8% net response rate yielding 203 surveys was achieved. The market structure-conduct-performance (SCP) paradigm from industrial organization provided a conceptual basis for testing a causal market definition model via LISREL. A latent exogenous variable (competitive intensity) and four latent endogenous variables (marketing orientation, technological orientation, market definition criteria, and market definition success) were used to develop and test hypothesized relationships among constructs. Research questions relating to market redefinition, market definition characteristics, and internal (within the firm) and external (competitive) market definition were also investigated. Market definition success was found to be positively associated with a marketing orientation and the use of market definition criteria. Technological orientation was not significantly related to market definition success. Customer needs were the key market definition characteristic to high-tech firms (technology, competition, customer groups, and products were also important). Market redefinition based on changing customer needs was the most effective of seven strategies tested. A majority of firms regularly defined their market at the corporate and product-line level within the firm. From a competitive perspective, industry, industry sector, and product-market definitions were used most frequently.

A planning model for needs analysis for subacute care

The purpose of this study is to produce a model to be used by state regulating agencies to assess demand for subacute care. In accomplishing this goal, the study refines the definition of subacute care, demonstrates a method for bed need assessment, and measures the effectiveness of this new level of care. This was the largest study of subacute care to date. Research focused on 19 subacute units in 16 states, each of which provides high-intensity rehabilitative and/or restorative care carried out in a high-tech unit. Each of the facilities was based in a nursing home, but utilized separate staff, equipment, and services. Because these facilities are under local control, it was possible to study regional differences in subacute care demand.^ Using this data, a model for predicting demand for subacute care services was created, building on earlier models submitted by John Whitman for the American Hospital Association and Robin E. MacStravic. The Broderick model uses the "bootstrapping" method and takes advantage of high technology: computers and software, databases in business and government, publicly available databases from providers or commercial vendors, professional organizations, and other information sources. Using newly available sources of information, this new model addresses the problems and needs of health care planners as they approach the challenges of the 21st century. ^

The development of direct ultra-fast PCR for forensic genotyping using short channel microfluidic systems with enhanced sieving matrices

There are situations in which it is very important to quickly and positively identify an individual. Examples include suspects detained in the neighborhood of a bombing or terrorist incident, individuals detained attempting to enter or leave the country, and victims of mass disasters. Systems utilized for these purposes must be fast, portable, and easy to maintain. The goal of this project was to develop an ultra fast, direct PCR method for forensic genotyping of oral swabs. The procedure developed eliminates the need for cellular digestion and extraction of the sample by performing those steps in the PCR tube itself. Then, special high-speed polymerases are added which are capable of amplifying a newly developed 7 loci multiplex in under 16 minutes. Following the amplification, a postage stamp sized microfluidic device equipped with specially designed entangled polymer separation matrix, yields a complete genotype in 80 seconds. The entire process is rapid and reliable, reducing the time from sample to genotype from 1-2 days to under 20 minutes. Operation requires minimal equipment and can be easily performed with a small high-speed thermal-cycler, reagents, and a microfluidic device with a laptop. The system was optimized and validated using a number of test parameters and a small test population. The overall precision was better than 0.17 bp and provided a power of discrimination greater than 1 in 106. The small footprint, and ease of use will permit this system to be an effective tool to quickly screen and identify individuals detained at ports of entry, police stations and remote locations. The system is robust, portable and demonstrates to the forensic community a simple solution to the problem of rapid determination of genetic identity.

Building Regional Security: Cooperation in the 21st Century: The Case of the Caribbean Regional Security System [RSS]

Small states that lack capacity and act on their own may fall victim to international and domestic terrorism, transnational organized crime or criminal gangs. The critical issue is not whether small Caribbean states should cooperate in meeting security challenges, but it is rather in what manner, and by which mechanisms can they overcome obstacles in the way of cooperation. The remit of the Regional Security System (RSS) has expanded dramatically, but its capabilities have improved very slowly. The member governments of the RSS are reluctant to develop military capacity beyond current levels since they see economic and social development and disaster relief as priorities, requiring little investment in military hardware. The RSS depends on international donors such as the USA, Canada, Great Britain, and increasingly China to fund training programs, maintain equipment and acquire material. In the view of most analysts, an expanded regional arrangement based on an RSS nucleus is not likely in the foreseeable future. Regional political consensus remains elusive and the predominance of national interests over regional considerations continues to serve as an obstacle to any CARICOM wide regional defense mechanism. Countries in the Caribbean, including the members of the RSS, have to become more responsible for their own security from their own resources. While larger CARICOM economies can do this, it would be difficult for most OECS members of the RSS to do the same. The CARICOM region including the RSS member countries, have undertaken direct regional initiatives in security collaboration. Implementation of the recommendations of the Regional Task Force on Crime and Security (RTFCS) and the structure and mechanisms created for the staging of the Cricket World Cup (CWC 2007) resulted in unprecedented levels of cooperation and permanent legacy institutions for the regional security toolbox. The most important tier of security relationships for the region is the United States and particularly USSOUTHCOM. The Caribbean Basin Security Initiative [CBSI] in which the countries of the RSS participate is a useful U.S. sponsored tool to strengthen the capabilities of the Caribbean countries and promote regional ownership of security initiatives. Future developments under discussion by policy makers in the Caribbean security environment include the granting of law enforcement authority to the military, the formation of a single OECS Police Force, and the creation of a single judicial and law enforcement space. The RSS must continue to work with its CARICOM partners, as well as with the traditional “Atlantic Powers” particularly Canada, the United States and the United Kingdom to implement a general framework for regional security collaboration. Regional security cooperation should embrace wider traditional and non-traditional elements of security appropriate to the 21st century. Security cooperation must utilize to the maximum the best available institutions, mechanisms, techniques and procedures already available in the region. The objective should not be the creation of new agencies but rather the generation of new resources to take effective operations to higher cumulative levels. Security and non-security tools should be combined for both strategic and operational purposes. Regional, hemispheric, and global implications of tactical and operational actions must be understood and appreciated by the forces of the RSS member states. The structure and mechanisms, created for the staging of Cricket World Cup 2007 should remain as legacy institutions and a toolbox for improving regional security cooperation in the Caribbean. RSS collaboration should build on the process of operational level synergies with traditional military partners. In this context, the United States must be a true partner with shared interests, and with the ability to work unobtrusively in a nationalistic environment. Withdrawal of U.S. support for the RSS is not an option.

A Look at Restaurant Commissaries

Restaurant commissaries range the full spectrum from simple storage of food and supplies to multi-million-dollar processing plants. The author discusses the cost effectiveness of commissary units, including their operating costs, quality control, and scope.

A comprehensive assessment of self -report experiences of adoptive parents of special needs children in Florida

Adoption of special needs children is now seen as a life long event whereby the adoptive child and family have unique needs. The need for postplacement resources throughout the life cycle of the adoption process is evident. This exploratory-descriptive research employed a random stratified cross-sectional design. The purpose of the study was to describe, identify, examine, and assess the relative influence of identified empirically and conceptually relevant variables of self-report experiences of adoptive parents of special needs children. Primary areas of exploration included: (1) adoptive children and families' characteristics, (2) postplacement service needs, utilization and satisfaction, and (3) adoptive parents' perceptions of their adoption experiences. A proportionate stratified random mail survey was used to obtain 474 families who had adopted special needs children from the 15 geographic districts which make up the state adoption social service agency in Florida. A 144-item survey questionnaire was used to collect basic information on demographic data, service provision, and adoption experiences. Four research questions were analyzed to test the effect the predictor variables had on willingness to adopt another special needs child, successful adoption, satisfying experience, and realism about problems. All four research questions revealed that the full model and the child's antecedent and the adoptive parents' intervening variable blocks were significant in explaining the variance in the dependent variables. The child's intervening variables alone were only significant in explaining the variance for one of the dependent variables. The results of the statistical analysis on the fifth research question and the three hypotheses determined that (1) only one postplacement service, crisis intervention, had a statistically significant impact on the adoptive parents' perceived level of satisfaction with the adoption experience; (2) adoptive parents who rate their adoption as successful are more likely to express a desire to adopt another special needs child; (3) the more adequate information on the child the adoptive parents perceived that they had prior to placement, the more they perceived they were realistic about the problems they would encounter; and (4) six specific postplacement services were found to be significant in predicting successful adoptions--crisis intervention, outpatient drug/alcohol treatment, maintenance subsidy, physical therapy, special medical equipment, and family counseling. Implications for the social work field and future research are discussed. ^

A planning model for needs analysis for subacute care

The purpose of this study is to produce a model to be used by state regulating agencies to assess demand for subacute care. In accomplishing this goal, the study refines the definition of subacute care, demonstrates a method for bed need assessment, and measures the effectiveness of this new level of care. This was the largest study of subacute care to date. Research focused on 19 subacute units in 16 states, each of which provides high-intensity rehabilitative and/or restorative care carried out in a high-tech unit. Each of the facilities was based in a nursing home, but utilized separate staff, equipment, and services. Because these facilities are under local control, it was possible to study regional differences in subacute care demand. Using this data, a model for predicting demand for subacute care services was created, building on earlier models submitted by John Whitman for the American Hospital Association and Robin E. MacStravic. The Broderick model uses the "bootstrapping" method and takes advantage of high technology: computers and software, databases in business and government, publicly available databases from providers or commercial vendors, professional organizations, and other information sources. Using newly available sources of information, this new model addresses the problems and needs of health care planners as they approach the challenges of the 21st century.

The Other Face of Cuban Medical Diplomacy: Domestic Impact

Since 1963 Cuba has provided medical assistance to third world countries while gaining international, political and economic support from its participating liaisons. But what exactly have been Cuba’s domestic consequences of such medical diplomacy? While the Cuban government sends many of its medical professionals and supplies abroad, the country suffers from extreme scarcity and a deterioration of its healthcare system. The purpose of my research is to enquire more on the consequences of such medical diplomacy on the Cuban healthcare system and how it has affected domestic medical infrastructure, health professionals working on the island as well as the quality of service. In carrying out this examination, I will rely on the use of information from books written by Cuban medical professionals on their personal experiences within the medical system, and patients treated on the island. I will also make active use of academic journals and articles on the Cuban healthcare system. Most of what has been written on Cuban medical diplomacy and internationalist missions are favorable critiques praising the Cuban government. Nevertheless, personal testimonies found in Dr. Dessy Mendoza Rivero’s Dengue: La Epidemia Secreta de Fidel Castro and in Dr. Jose Luis Comas and Dr. Luis Ovidio Gonzales’ Cuba: Medicina y Revolucion reveal a different side to this seemingly pleasant and good natured international exchange. As a Cuban who lived on the island, I personally suffered the consequences of such medical diplomacy and believe others will find benefit in arming themselves with knowledge on the issue.

Atuação dos profissionais no atendimento às famílias nos centros de atenção psicossocial

This study aimed to evaluate the work of professionals to care for families in Psychosocial Care Centers ( C APS) of Rio Grande do Norte ( RN), from the roles and functions performed by these professional services. For this, it was pointed out the following objectives: To describe the profile and the activities conducted by mental health teams in the RN CAPS ; Know the opinion of professionals in the mental health teams of the poli ti c , practices and training in mental health; Check the suitability of the roles and functions of professionals working in the RN CAPS in relation to care for families . This is an analytic al cross - sectional study of quantitative and qualitative approach . Data were c ollected through a questionnaire in 33 CAPS RN, between March and October 2014 , after being approved by the Research Ethics Committee / UFRN , opinion nº217.808 , CAAE : 10650612.8. 1001.5537 , on March 1 2013. T he sample was adopted , defined by inclusion and exclusion criteria , and is composed of 183 professionals. The database preparation followed two steps: 1. Preparation and processing of data of closed questions of the questionnai re concerning the characterization and practices in mental health research subjects through informational resource Statistical Package for Social Scienses (SPSS) Statistics version 20.0 ; 2. To check the significance level was chosen by applying the chi - squ are test. Preparation and treatment of the corpus formed by the answers to open questions relating to the policies, practices and training in psychiatry through Analyse lexicale pair Contexte software d' un Ensemble of Segments of Texte ( ALCESTE) together a nd categorized by content analysis technique , Bardin (2004) . The data analysis is supported in the literature . It m ade explicit the results through three articles waxing the following results. In the first, participants profile was characterized by a predo minance of females (76.5 %), aged 40 - 58 years ( 61.7 %). They work between 30 and 40 hours per week (63.5 %), working in mental health for over 10 years ( 98.4%). The sample directs the care of family groups ( 65.7%), predominantly the care team of social worke rs, nurses, psychologists and occupational therapists . The doctor performs emergency care without interaction with the staff (48.6%) . On the difficulties encountered in services are ranked in : materials and supplies ( 75.1%), financial ( 78.5%) and structura l ( 66.9%). The second article contains qualitative data organized into five categories : Promoting the rehabilitation of users of CAPS ; Needs training ; Conflicts and satisfactions of teamwork ; Practices developed in CAPS ; Effective difficulties of Mental He alth Policy . The third article highlights the inadequacy of care for families ( 93.4%) and comparing the care families and groups in CAPS both types show to be inadequate : family ( 92.63%), groups ( 92, 60%). The main data obtained reveal the urgent need for transformation in psychosocial care . It shows also the importance of investments in inputs, physical structure and training of human resources for the CAPS.

Análise computacional de imagens de lâminas delgadas na predição dos parâmetros hidráulicos porosidade e condutividade hidráulica do aquífero Barreiras-RN

The understanding of the occurrence and flow of groundwater in the subsurface is of fundamental importance in the exploitation of water, just like knowledge of all associated hydrogeological context. These factors are primarily controlled by geometry of a certain pore system, given the nature of sedimentary aquifers. Thus, the microstructural characterization, as the interconnectivity of the system, it is essential to know the macro properties porosity and permeability of reservoir rock, in which can be done on a statistical characterization by twodimensional analysis. The latter is being held on a computing platform, using image thin sections of reservoir rock, allowing the prediction of the properties effective porosity and hydraulic conductivity. For Barreiras Aquifer to obtain such parameters derived primarily from the interpretation of tests of aquifers, a practice that usually involves a fairly complex logistics in terms of equipment and personnel required in addition to high cost of operation. Thus, the analysis and digital image processing is presented as an alternative tool for the characterization of hydraulic parameters, showing up as a practical and inexpensive method. This methodology is based on a flowchart work involving sampling, preparation of thin sections and their respective images, segmentation and geometric characterization, three-dimensional reconstruction and flow simulation. In this research, computational image analysis of thin sections of rocks has shown that aquifer storage coefficients ranging from 0,035 to 0,12 with an average of 0,076, while its hydrogeological substrate (associated with the top of the carbonate sequence outcropping not region) presents effective porosities of the order of 2%. For the transport regime, it is evidenced that the methodology presents results below of those found in the bibliographic data relating to hydraulic conductivity, mean values of 1,04 x10-6 m/s, with fluctuations between 2,94 x10-6 m/s and 3,61x10-8 m/s, probably due to the larger scale study and the heterogeneity of the medium studied.

Análise de confiabilidade de um moto-ventilador centrífugo: aplicação no sistema de pressurização de subestação industrial para garantia da segurança operacional na indústria petroquímica

The need of the oil industry to ensure the safety of the facilities, employees and the environment, not to mention the search for maximum efficiency of its facilities, makes it seeks to achieve a high level of excellence in all stages of its production processes in order to obtain the required quality of the final product. Know the reliability of equipment and what it stands for a system is of fundamental importance for ensuring the operational safety. The reliability analysis technique has been increasingly applied in the oil industry as fault prediction tool and undesirable events that can affect business continuity. It is an applied scientific methodology that involves knowledge in engineering and statistics to meet and or analyze the performance of components, equipment and systems in order to ensure that they perform their function without fail, for a period of time and under a specific condition. The results of reliability analyzes help in making decisions about the best maintenance strategy of petrochemical plants. Reliability analysis was applied on equipment (bike-centrifugal fan) between the period 2010-2014 at the Polo Petrobras Guamaré Industrial, situated in rural Guamaré municipality in the state of Rio Grande do Norte, where he collected data field, analyzed historical equipment and observing the behavior of faults and their impacts. The data were processed in commercial software reliability ReliaSoft BlockSim 9. The results were compared with a study conducted by the experts in the field in order to get the best maintenance strategy for the studied system. With the results obtained from the reliability analysis tools was possible to determine the availability of the centrifugal motor-fan and what will be its impact on the security of process units if it will fail. A new maintenance strategy was established to improve the reliability, availability, maintainability and decreased likelihood of Moto-Centrifugal Fan failures, it is a series of actions to promote the increased system reliability and consequent increase in cycle life of the asset. Thus, this strategy sets out preventive measures to reduce the probability of failure and mitigating aimed at minimizing the consequences.