Abstract Genetic analyses through molecular markers always depend on efficient protocols for DNA extraction and standardized PCR conditions. Random Amplified Polymorphic DNA (RAPD) is an inexpensive protocol that can provide information about the genetic diversity among Capsicum accessions within a germplasm bank, but it is highly sensitive to reaction conditions and manipulation. This work presents an optimized protocol for DNA isolation from dry Capsicum seeds, and a standardization of five conditions for PCR using RAPD primers. The RAPD protocol was optimized for the following parameters: 50 ng of DNA template per mix, 2.5 mM MgCl2, 200 µM dNTP's, 25 ng of primer per mix, and 45 thermal cycles, resulting in reproducible and clear amplified fragments. Good quality products were presented in the extractions and amplifications carried out; therefore, the optimized protocols are sufficient to be used in future work with RAPD. (RAPD bank manipulation seeds primers parameters 5 mix 2 2. MgCl2 MgCl 20 dNTPs, dNTPs dNTP s, s dNTP's 4 cycles fragments out therefore

The impact of school closings on reducing the number of cases of influenza-like illness during an outbreak of influenza A (H1N1), which reached pandemic proportions, was assessed, along with other control measures, in the two main cities of Tierra del Fuego Province in southern Argentina. The incidence before and after the school closings in 2009 was compared by means of the t-test for related samples. By week 40, 6 901 cases of influenza-like illness had been detected, 281 of which were confirmed as influenza A (H1N1) through laboratory tests; 38 patients were hospitalized. After the intervention, there were nearly 10 times fewer cases than the average recorded in the health centers. The results indicate that closing schools during the influenza A (H1N1) outbreak resulted in a significantly lower incidence of influenza-like illness. However, the impact of other measures, such as case management and protection against exposure, should not be ignored. Timely implementation of this intervention, together with other measures, can help minimize the spread of influenza outbreaks.

Se evaluó el efecto del cierre de las escuelas en la reducción del número de casos de enfermedades del tipo influenza durante un brote de influenza A por el virus H1N1, de características pandémicas, en las dos principales ciudades de la provincia de Tierra del Fuego, en la región austral de Argentina, en conjunto con otras medidas de control. Se comparó la incidencia antes y después del cierre de las escuelas en 2009 mediante la prueba de la t para muestras relacionadas. Hasta la semana 40 se detectaron 6 901 casos de enfermedades del tipo influenza, de ellos 281 se confirmaron como influenza A H1N1 mediante pruebas de laboratorio; 38 pacientes recibieron hospitalización. Se observó una reducción de cerca de 10 veces en la incidencia promedio registrada en los centros de salud después de la intervención. Los resultados indican que el cierre de las escuelas durante el brote de influenza A H1N1 se acompañó de una reducción significativa de la incidencia de enfermedades tipo influenza, aunque no se debe descartar el efecto de otras acciones, como el tratamiento de los casos y la profilaxis de sus contactos. Esta intervención, aplicada oportunamente, junto con otras medidas, puede contribuir a reducir la propagación de los brotes de influenza."

The genus Arachis (Fabaceae) native to South America, contains 80 species divided into nine sections, three of which contain species of special economic importance such as the cultivated peanut (Arachis hypogaea), belonging to the section Arachis, and some perennial forage species from sections Caulorrhizae and Rhizomatosae. We used microsatellite markers to assay genetic variability among 77 accessions of four species from section Rhizomatosae, the diploid Arachis burkartii (2n = 2x = 20) and the tetraploid Arachis glabrata, Arachis pseudovillosa and Arachis nitida (2n = 4x = 40). A total of 249 alleles were found in the fifteen loci analyzed and a high degree of intra and interspecific polymorphism was detected. The lowest intraspecific variation occurred in Arachis burkartii, while the smallest estimated interspecific value was between A. nitida and A. pseudovillosa and the largest was between A. burkartii and A. nitida. High observed heterozygosity was detected in A. glabrata. The diploid accessions grouped in one cluster and the tetraploid accessions in another. It was possible to distinguish all 77 accessions and the genetic distance between accessions could not be correlated with geographic origin.

Nearly 65,000 citrus EST (Expressed Sequence Tags) have been investigated using the CitEST project database. Microsatellites were investigated in the unigene sequences from Citrus spp. and Poncirus trifoliata. From these sequences, approximately 35% of the non-redundant ESTs contained SSRs. The frequencies of different SSR motifs were similar between Citrus spp and trifoliate orange. In general, mononucleotide repeats appeared to be the most abundant SSRs in the CitEST database, but we also identify di-, tri-, tetra-, penta- and hexanucleotide repeats. The AG/CT and AAG/CTT were the most common dinucleotide and trinucleotide motifs, with frequencies of 54.4% and 25.2%, respectively. Primer sequences flanking SSR motifs were successfully designed and synthesized. After in silico polymorphism analysis, a subset of sixty-eight primers was validated in different Citrus spp. and Poncirus trifoliata. PCR-amplification revealed polymorphism in citrus with all tested primer pairs and showed the potential of these markers for linkage mapping. Our study showed that the CitEST database can be exploited for the development of SSR markers that can amplify Citrus spp. and related genus for comparative mapping and other genetic analyses.