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.

Multiplex PCR for Assessment of Tetracycline Resistance (tet) Genes in Antibiotic-Resistant Soil Bacteria and the Ecological Implications

Antibiotics are becoming increasingly prevalent in bacterial communities due to clinical and agricultural misuse and overuse in their environment. As exposure increases, so does the incidence of microbial resistance. Such is the case with bacterial resistance to tetracyclines, a phenotype often acquired through the horizontal gene transfer of tet genes between bacteria. The objective of this project was to analyze the bacterial diversity of tet resistance genes in soil from Miami-Dade County. Bacterial isolates were Gram-stained and the Kirby-Bauer antibiotic disk diffusion test was performed to determine each bacterium’s degree of resistance. The 16S rRNA gene from antibiotic-resistant isolates was amplified by PCR and sequenced to identify the isolates. All isolates’ tet genes were amplified by multiplex PCR, sequenced, and compared. Among eight isolates, three distinct species were positively identified based on their 16S rRNA sequences and four distinct tet genes were identified, though all tested susceptible to tetracycline via the Kirby-Bauer test. This project clarifies some aspects of the ecology of antibiotic resistance genes, their natural ecological function and the potential for the expansion of intrinsic multi-antibiotic resistance into new ecosystems and/or hosts.

Determinación de factor reumatoide, pcr y vsg en cincuenta pacientes con artritis reumatoide que reciben terapia biológica en el Hospital José Carrasco Arteaga de la ciudad de Cuenca

La Terapia biológica aplicada a pacientes diagnosticados de Artritis Reumatoide ha demostrado tener buenos resultados paraclínicos en estudios realizados en otros países. Ejecutamos esta investigación para conocer mediante Laboratorio (PCR-VSG y FR), si dichos cambios se presentan en pacientes con AR que reciben tratamiento biológico en nuestro medio. Métodos: Se realizó un estudio cuasiexperimental, seleccionando al azar 50 pacientes con diagnóstico de Artritis Reumatoide sin enfermedad concomitante, que reciben tratamiento biológico en el Hospital José Carrasco de la Ciudad de Cuenca, se revisó en las Historias Clínicas los valores preterapéuticos de PCR, VSG y FR, luego se realizaron controles a partir de los 5 meses de inicio de la terapia biológica y se confrontaron resultados. El análisis incluyó Chi cuadrado, Valor p, Desvio Estándar y diferencia de medias. Resultados: De los 50 pacientes estudiados, el PCR fue positivo en el 100de pacientes antes y después de la Terapia biológica. Los valores de VSG fueron positivos preterapia en el 70y post-terapia en el 40, (p= 0.003). El FR fue positivo en el 84antes del tratamiento, luego del mismo el porcentaje de FR positivo fue 60, (p= 0.008). La media de los valores de PCR antes del tratamiento fue 12.8ñ14.2DE y luego del tratamiento 5.6 ñ 6.5DE (p= 0.002) la media para la VSG preterapia fue 33.4 ñ 21.3DE y post-terapia 17.2 ñ 6.7DE (p= 0,0001). Para el FR la media antes de la terapia fue 120.6 ñ 97.9 y luego de la terapia 100.2ñ 122.3DE (p= 0.360). Conclusiones: Los valores de PCR, VSG y FR de cincuenta pacientes con Artritis Reumatoide, presentaron un descenso significativo, luego de recibir tratamiento biológico en el Hospital José Carrasco Arteaga de la Ciudad de Cuenca

The Coming of Post-reflexive Society: Commodification and Language in Digital Capitalism

Language is a unique aspect of human communication because it can be used to discuss itself in its own terms. For this reason, human societies potentially have superior capacities of co-ordination, reflexive self-correction, and innovation than other animal, physical or cybernetic systems. However, this analysis also reveals that language is interconnected with the economically and technologically mediated social sphere and hence is vulnerable to abstraction, objectification, reification, and therefore ideology – all of which are antithetical to its reflexive function, whilst paradoxically being a fundamental part of it. In particular, in capitalism, language is increasingly commodified within the social domains created and affected by ubiquitous communication technologies. The advent of the so-called ‘knowledge economy’ implicates exchangeable forms of thought (language) as the fundamental commodities of this emerging system. The historical point at which a ‘knowledge economy’ emerges, then, is the critical point at which thought itself becomes a commodified ‘thing’, and language becomes its “objective” means of exchange. However, the processes by which such commodification and objectification occurs obscures the unique social relations within which these language commodities are produced. The latest economic phase of capitalism – the knowledge economy – and the obfuscating trajectory which accompanies it, we argue, is destroying the reflexive capacity of language particularly through the process of commodification. This can be seen in that the language practices that have emerged in conjunction with digital technologies are increasingly non-reflexive and therefore less capable of self-critical, conscious change.