Testing and Validation of High Density Resequencing Microarray for Broad Range Biothreat Agents Detection

Rapid and effective detection and identification of emerging microbiological threats and potential biowarfare agents is very challenging when using traditional culture-based methods. Contemporary molecular techniques, relying upon reverse transcription and/or polymerase chain reaction (RT-PCR/PCR) provide a rapid and effective alternative, however, such assays are generally designed and optimized to detect only a limited number of targets, and seldom are capable of differentiation among variants of detected targets. To meet these challenges, we have designed a broad-range resequencing pathogen microarray (RPM) for detection of tropical and emerging infectious agents (TEI) including biothreat agents: RPM-TEI v 1.0 (RPM-TEI). The scope of the RPM-TEI assay enables detection and differential identification of 84 types of pathogens and 13 toxin genes, including most of the class A, B and C select agents as defined by the Centers for Disease Control and Prevention (CDC, Atlanta, GA). Due to the high risks associated with handling these particular target pathogens, the sensitivity validation of the RPM-TEI has been performed using an innovative approach, in which synthetic DNA fragments are used as templates for testing the assay's limit of detection (LOD). Assay specificity and sensitivity was subsequently confirmed by testing with full-length genomic nucleic acids of selected agents. The LOD for a majority of the agents detected by RPM-TEI was determined to be at least 104 copies per test. Our results also show that the RPM-TEI assay not only detects and identifies agents, but is also able to differentiate near neighbors of the same agent types, such as closely related strains of filoviruses of the Ebola Zaire group, or the Machupo and Lassa arenaviruses. Furthermore, each RPM-TEI assay results in specimen-specific agent gene sequence information that can be used to assess pathogenicity, mutations, and virulence markers, results that are not generally available from multiplexed RT-PCR/PCR-based detection assays

Transformation and analysis of tobacco plant var Petit havana with T-urf13 gene under anther-specific TA29 promoter

T-urf13, a well-documented cms-associated gene from maize, has been shown to render methomyl sensitivity to heterologous systems like rice, yeast and bacteria when expressed constitutively. Since these transgenic plants were fertile, it was hypothesized that T-urf13 gene if expressed in anthers may result in male sterility that could be used for hybrid seed production. Hence, this work was aimed at analysing whether T-urf13 gene when expressed in anthers can result in male sterile plants or requires methomyl treatment to cause male sterility (controllable). This is the first report of transformation of tobacco with T-urf13 gene under anther-specific promoter (TA29) with or without mitochondrial targeting sequence. Most of the transgenic plants obtained were fertile; this was surprising as many male sterile plants were expected as T-urf13 gene is a cms associated gene. Our results suggest that it may not be possible to obtain male sterility by expressing URF13 in the anther by itself or by methomyl application

Adherence to a Mediterranean diet in Morocco and its correlates: cross-sectional analysis of a sample of the adult Moroccan population

<p>Abstract</p> <p>Background</p> <p>Dietary habits in Morocco are changing and the causes are not well understood. This study aimed to analyse socio-demographic factors associated with adherence to the Mediterranean diet (MeDi) in a national random sample of the adult Moroccan population.</p> <p>Methods</p> <p>The data collected in this cross-sectional survey included socio-demographic factors and a food frequency questionnaire. MeDi adherence was assessed in 2214 individuals with complete dietary data. MeDi adherence was measured according to a simplified MeDi score based on the weekly frequency of intake of eight food groups (vegetables, legumes, fruits, cereal or potatoes, fish, red meat, dairy products and olive oil) with the use of the sex specific medians of the sample as cut-offs. A value of 0 or 1 was assigned to consumption of each component according to its presumed detrimental or beneficial effect on health. Logistic regression was used to estimate the association between MeDi adherence (low score 1-4 vs. high 5-8) and other factors.</p> <p>Results</p> <p>Mean age of the sample was 41.4 (standard deviation 15.3) years, 45.4% were men and 29.9% had a low MeDi adherence. Married subjects (adjusted odds ratio ORa=0.68, 95% CI 0.55-0.84) were less likely to have a low MeDi adherence compared to single, divorced or widowed persons. Persons from rural areas (ORa=1.46, 95% CI: 1.02-2.08), were more often low MeDi adherents compared to those from urban areas. Obese persons (ORa=1.56, 95% CI: 1.16-2.11) were more prone to low MeDi adherence than normal weight individuals.</p> <p>Conclusion</p> <p>MeDi is far from being a universal pattern in the Moroccan population. Intervention strategies should be implemented in target groups to maintain the traditional MeDi pattern considered as the original diet in Morocco.</p

Imaging in retroperitoneal soft tissue sarcoma.

Patients with retroperitoneal sarcoma can present to a variety of clinicians with non-specific symptoms and retroperitoneal sarcomas can be incidental findings. Failure to recognize retroperitoneal sarcomas on imaging can lead to inappropriate management in non-specialist centers. Therefore it is critical that the possibility of retroperitoneal sarcoma should be considered with prompt referral to a soft tissue sarcoma unit. This review guides clinicians through a diagnostic pathway, introduces concepts in response assessment and new imaging developments