Development of real-time PCR method for the detection and the quantification of a new endogenous reference gene in sugar beet "Beta vulgaris L.": GMO application.

KEY MESSAGE : Here, we describe a new developed quantitative real-time PCR method for the detection and quantification of a new specific endogenous reference gene used in GMO analysis. The key requirement of this study was the identification of a new reference gene used for the differentiation of the four genomic sections of the sugar beet (Beta vulgaris L.) (Beta, Corrollinae, Nanae and Procumbentes) suitable for quantification of genetically modified sugar beet. A specific qualitative polymerase chain reaction (PCR) assay was designed to detect the sugar beet amplifying a region of the adenylate transporter (ant) gene only from the species of the genomic section I of the genus Beta (cultivated and wild relatives) and showing negative PCR results for 7 species of the 3 other sections, 8 related species and 20 non-sugar beet plants. The sensitivity of the assay was 15 haploid genome copies (HGC). A quantitative real-time polymerase chain reaction (QRT-PCR) assay was also performed, having high linearity (R (2) > 0.994) over sugar beet standard concentrations ranging from 20,000 to 10 HGC of the sugar beet DNA per PCR. The QRT-PCR assay described in this study was specific and more sensitive for sugar beet quantification compared to the validated test previously reported in the European Reference Laboratory. This assay is suitable for GMO quantification in routine analysis from a wide variety of matrices.

Variation of volatiles in Tunisian populations of Thymbra capitata (L.) Cav. (Lamiaceae).

Volatiles from 14 wild Tunisian populations of Thymbra capitata (=Thymus capitatus Hoffmanns. et Link=Coridothymus capitatus Rchb.f.), sampled in five ecological areas (sub-humid, upper semi-arid, mean semi-arid, lower semi-arid, and upper arid areas) were analyzed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS) techniques. Thirty-nine constituents representing 94.2 to 99.5% of the total oil were identified. Carvacrol (38; 51.1-75.9%), p-cymene (13; 3.7-15%), γ-terpinene (12; 1.4-11.9%), and trans-ß-caryophyllene (22; 2.9-4.6%) are the major compounds. A significant variation among populations and population's bioclimatic stage for the majority of compounds was shown. The chemical population structure, estimated using a principal-component analysis (PCA) and an unweighted pair group method with arithmetic mean (UPGMA) cluster analysis performed on all populations and compounds and based on Euclidean distances among populations, was high. Both methods allowed separation of the populations into distinct groups defined rather by minor than by major compounds. The spatial compound distribution is linked to ecological factors, indicating that local selective environmental factors influence chemical-composition diversity. Conservation strategies should involve all populations because of their low size and their high level of destruction. Populations exhibiting particular compounds other than the major ones should first be protected. In situ conservation of populations should be accomplished appropriately according to bioclimate.

Neonatal cardiogenic shock revealing obstructive cardiac Hibernoma: case report.

BACKGROUND: Cardiac Hibernomas are very rare benign tumors and usually remain asymptomatic. Neonatal cardiogenic shock due to cardiac tumors is extremely very rare. Until this date a few cases of cardiac hibernoma have been reported in the literature. Transthoracic echocardiography help in the differential diagnosis, but the definitive diagnosis is histological. The management strategy is not clearly codified. The Aim is to report and discuss the clinical features of a cardiac Hibernoma and review the relevant literature. CASE PRESENTATION: We describe a case of a 2-day-old Caucasian full-term male neonate admitted in neonate intensive care with cardiogenic shock, having fluid resuscitation and inotropic drugs. Ventilatory support was started immediately with the subsequent reestablishment of normal blood pressure. Then he was transferred to the echocardiography laboratory. Transthoracic echocardiography showed two echogenic masses in the right atrium and right ventricle. The masses were extended to the pulmonary trunk. Pulmonary artery flow measurements showed the presence of pulmonary and tricuspid obstruction. Surgery was rapidly considered since the baby was hemodynamically unstable. Intraoperative evaluation showed a mass embedded in the interventricular septum that occupy the right ventricular cavity and the right atrium. The tumor involved also the chordae of the tricuspid. Partial resection was done. Tricuspid valve repair was performed by construction of new chordae from the autologous pericardium. The specimen was sent for histopathological analysis. The baby died immediately after surgery. Histological examination of the surgical specimen revealed clear multivacuolated cells filled with lipid droplets and granular intense eosinophilic cytoplasm which confirms the diagnosis of Hibernoma. CONCLUSION: Cardiac Hibernomas are rare benign tumors. The prognosis and treatment strategy is closely dependent on the location, initial clinical presentation and possible complications. The prognosis can be unfavorable if the tumor was obstructive and infiltrate the myocardium.

Variation of the chemical composition of essential oils in Tunisian populations of Thymus algeriensis Boiss. et Reut. (Lamiaceae) and implication for conservation.

The variation of the essential-oil composition among 14 Tunisian natural populations of Thymus algeriensis Boiss. et Reut. (=Thymus hirtus Willd. ssp. algeriensis Boiss. et Reut.) was assessed by GC (RI) and GC/MS. The populations were collected from different geographical regions belonging to the sub-humid, upper semi-arid, mean semi-arid, lower semi-arid, and upper-arid bioclimates. A total of 47 constituents, representing 81.0 to 96.5% of the total oil, were identified. The main volatiles at the species level were 1,8-cineole (17.7%), alpha-pinene (15.5%), and camphor (8.2%). A high variation among populations for the majority of the compounds was shown. Camphor (0.2-14.0%), linalool (0.2-22.4%), borneol (<0.01-24.3%), caryophyllene oxide (<0.01-18.8%), thymol (<0.01-54.9%), gamma-terpinene (0.4-6.5%), alpha-copaene (0.4-7.6%), linalyl acetate (<0.01-6.4%), and methyl eugenol (<0.01-6.9%) were the main constituents differentiating the populations. The chemical differentiation among populations, assessed by principal component analysis (PCA) and a UPGMA (unweighted pair-group method with averaging) cluster analysis performed on all populations and compounds, was high. Six chemotypes according to the main compounds have been distinguished, i.e., caryophyllene oxide/1,8-cineole/alpha-pinene, 1,8-cineole/alpha-pinene, 1,8-cineole/alpha-pinene/camphor, borneol/1,8-cineole/alpha-pinene, linalool, and thymol chemotypes. The spatial chemotype distribution was linked to the geographic distance among populations rather than to bioclimates, indicating that local selective environmental factors act on the chemotype diversity. The high chemical variation among populations according to their geographical and bioclimatic distribution imposes that conservation strategies of populations should be made appropriately, taking into account these factors. The in situ and ex situ conservation strategies should concern all populations representing the different chemotypes.