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PubMed | Biomedica, Biosafety Unit, Princess Sumaya University for Technology, Northwestern University and Knowledge Sector; Royal Scientific Society; Amman
Type: Journal Article | Journal: GM crops & food | Year: 2014

The Cauliflower Mosaic Virus 35S promoter sequence, CaMV P-35S, is one of several commonly used genetic targets to detect genetically modified maize and is found in most GMOs. In this research we report the finding of an alternative P-35S sequence and its incidence in GM maize marketed in Jordan. The primer pair normally used to amplify a 123 bp DNA fragment of the CaMV P-35S promoter in GMOs also amplified a previously undetected alternative sequence of CaMV P-35S in GM maize samples which we term V3. The amplified V3 sequence comprises 386 base pairs and was not found in the standard wild-type maize, MON810 and MON 863 GM maize. The identified GM maize samples carrying the V3 sequence were found free of CaMV when compared with CaMV infected brown mustard sample. The data of sequence alignment analysis of the V3 genetic element showed 90% similarity with the matching P-35S sequence of the cauliflower mosaic virus isolate CabbB-JI and 99% similarity with matching P-35S sequences found in several binary plant vectors, of which the binary vector locus JQ693018 is one example. The current study showed an increase of 44% in the incidence of the identified 386 bp sequence in GM maize sold in Jordans markets during the period 2009 and 2012.

Mendes M.E.,University of Sao Paulo | Fagundes C.C.,Biomedica | Do Porto C.C.,Servico de Bioquimica Clinica | Bento L.C.,Biomedica | And 3 more authors.
Jornal Brasileiro de Patologia e Medicina Laboratorial | Year: 2011

Water is a reagent used in most laboratory tests and, therefore, must follow stringent quality control standards. The urban water supply has organic molecules, inorganic ions, particles, colloids, gases, bacteria and their products, which may alter laboratory test results and cause occasional errors and mechanical failures in diagnostic equipment. To remove these impurities, it is necessary to use a combination of purification technologies. There are several organizations that specify reagent water standards to minimize its interference in laboratory assays. Most laboratories set standards established by the Clinical and Laboratory Standards Institute (CLSI), which classifies the type of water as follows: clinical laboratory reagent water (CLRW), special reagent water (SRW) and instrumental feed water (IFW). The quality monitoring is performed by means of assessing the resistivity, conductivity, total organic carbon (TOC), microbial control and endotoxins. The parameters are evaluated in accordance with the frequency determined by the standard used. In this article we discuss the importance of water employed in laboratory procedures, its quality control and its interference in laboratory assays.

Schoppet M.,University of Marburg | Hofbauer L.C.,TU Dresden | Brinskelle-Schmal N.,Biomarker Design Forschung GmbH | Varennes A.,Central Biochemical Laboratory | And 5 more authors.
Journal of Clinical Endocrinology and Metabolism | Year: 2012

Context: Calcification inhibitor deficiencies, mineral imbalance, and phenotypic transformation of vascular cells to osteogenic cells initiate and sustain vascular calcification. Fibroblast growth factor- 23 (FGF23) is a key molecule regulating mineral homeostasis. Objective: Our objective was to assess the association of serum FGF23 levels with mineral metabolism parameters and abdominal aortic calcification (AAC) in men. Design: This was a cross-sectional analysis in the STRAMBO cohort. Setting: Men holding a private health insurance cover with Mutuelle de Travailleurs de la Région Lyonnaise were included in the study. Participants: Participants included male volunteers aged 20-87 (n = 1130). Interventions: Nonfasting blood collection was done. AAC was semiquantitatively assessed from vertebral fracture assessment scans obtained using dual-energy x-ray absorptiometry. Main Outcome Measures: We evaluated the association between FGF23 concentration and AAC severity in men. Results: In 350 men aged 60 yr or younger, FGF23 levels decreased with age (r = -0.21; P < 0.001) but were not associated with any other parameter. In 780 men aged over 60 yr, serum FGF23 correlated with age (r = 0.37; P < 0.001) and, after adjustment for confounders, with glomerular filtration rate (r = -0.31; P < 0.001) and PTH levels (r = 0.25; P < 0.001). After adjustment for confounders, self-reported ischemic heart disease, diabetes mellitus as well as higher concentrations of C-reactive protein and osteoprotegerin were all associated with higher FGF23 levels. After adjustment for confounders, subjects in the highest FGF23 quartile had higher prevalence of severe AAC compared with the three lower quartiles combined (odds ratio = 1.88; 95% confidence interval = 1.22-2.85; P < 0.005). Conclusions: In healthy older men, circulating FGF23 is associated with parameters of mineral metabolism, including bone metabolism-regulating cytokines, and with severe AAC independent of traditional risk factors. Copyright © 2012 by The Endocrine Society.

Tarasconi A.C.,University Of Passo Fundo | Silva E.V.,Biologa. Genesis Clinica de Reproducao Humana | Tarasconi B.V.,University Of Passo Fundo | Verzeletti F.,Biomedica | Tarasconi D.V.,University Of Passo Fundo
Jornal Brasileiro de Reproducao Assistida | Year: 2010

Objective: To develop a comparative study among intracytoplasmic morphologically selected sperm injection (IMSI) and conventional intracytoplasmic sperm injection (ICSI), assessing the efficacy of both in fertilization rates, cleavage and embryo quality. Methods: 36 cycles of intracytoplasmic sperm injection have been analyzed. In these cycles were obtained 603 mature oocytes (MII), which were injected with sperm selected by two different techniques. The oocytes obtained of each patient were divided into two groups (A and B). In group A (302 oocytes), were employed, to assess the morphology of sperm to be injected, the conventional equipment that provides optic magnification of 400 X (conventional ICSI). In group B (301 oocytes), were employed an equipment that provides an optic magnification of 6800X, (IMSI). After embryo culture was performed, fertilization rates, cleavage and embryo quality until day 3, on each group, were analyzed. Results: The total fertilization rate in group A was 78,81%, and in group B, 81,06%. The cleavage rate in group A was 97,29%, and in group B, 96,41%. The percentage of good quality embryos (grade I e II), in day 2, in group A was 79,53% and in group B, 78,13%. The percentage of good quality embryos on day 3, in group A was 77,41% and in group B 78,81%. Conclusion: There were no significant difference in fertilization rates, cleavage and embryo quality to day 3, comparing these two methods of sperm selection. Copyright - Todos os direitos reservados a SBRA - Sociedade Brasileira de Reprodução Assistida.

Piemonte S.,University of Rome La Sapienza | Romagnoli E.,University of Rome La Sapienza | Bratengeier C.,Biomarker Design Forschungs GmbH | Woloszczuk W.,Biomedica | And 4 more authors.
Journal of Endocrinological Investigation | Year: 2012

Objective: This study was carried out in order to evaluate the effect of 18-month treatment with PTH (1-34) or PTH (1-84) on serum sclerostin levels in humans. Subjects and methods: We investigated 10 women with severe osteoporosis, previously treated with alendronate and 20 untreated osteoporotic women. Subjects with severe osteoporosis were randomly divided into 2 groups of 5 patients each; the first group was treated with 20 μg of PTH (1-34) and the second one with 100 μg of PTH (1-84) according to an open-label design. Fasting blood samples were collected at baseline and at 2, 4, and 24 h after hormone administration. The same protocol was followed at month 1, 6, 12, 18. Serum sclerostin levels were measured at each time point by a sandwich-type enzyme-linked immunosorbent assay. Results: Basal serum sclerostin levels were not significantly different between patients previously treated with alendronate and those never treated. No significant acute change of serum sclerostin levels was observed after PTH administration. Fitting a mixed effect regression model, we found a significant time effect (p=0.0012) using the sclerostin level as the response variable and the month of drug administration as a single covariate. Treatment with both PTH molecules induced a monthly mean reduction of sclerostin levels of 0.1956 pmol/l. Conclusions: Our results indicate that long-term therapy with PTH (1-34) or PTH (1-84) in women with osteoporosis previously treated with alendronate is associated with a reduction in circulating sclerostin levels. This is a putative mechanism through which PTH performs its anabolic action. ©2012, Editrice Kurtis.

Szulc P.,University of Lyon | Hawa G.,BioMedica | Boutroy S.,University of Lyon | Vilayphiou N.,University of Lyon | And 4 more authors.
Journal of Clinical Endocrinology and Metabolism | Year: 2011

Context: Osteoprotegerin (OPG) is an inhibitor of bone resorption, but its relationship to bone microarchitecture remains unclear. Objective: Our objective was to study the relationship between OPG concentration and bone microarchitecture in men. Design, Setting, and Participants: We conducted a cross-sectional study of a population-based cohort of 1149 men aged 20-87 yr. Interventions: We assessed bone microarchitecture at the distal radius and tibia by high-resolution peripheral quantitative computed tomography (XtremeCT Scanco) and measured serum OPG concentration and bone turnover markers: osteocalcin, bone-specific alkaline phosphatase, N-terminal extension type I collagen propeptide, C-terminal type 1 collagen telopeptide, and urinary deoxypyridinoline. Main outcome measures: Differences were assessed in bone microarchitectural parameters across the OPG quartiles in the models adjusted for age, weight, height, physical activity, ischemic heart disease, diabetes mellitus, calcium intake, serum levels of free testosterone, bioavailable 17β-estradiol, PTH, 25-hydroxycholecalciferol, and creatinine. Results: After adjustment for the confounders, men in the highest (fourth) quartile of OPG levels (>4.55pmol/liter)hadhighertotal cross- sectionalareaandtrabecularareaatthedistalradiusanddistal tibia (3.3-6.0%, P < 0.05). At both skeletal sites, the highest OPG quartile was associated with lower cortical thickness (8.2%, P < 0.001, and 3.7%, P < 0.05) and volumetric bone mineral density (vBMD, 2.7%, P < 0.001, and 1.6%, P < 0.005) compared with the three lower quartiles combined. Associations of OPG level with trabecular vBMD, number, thickness, and distribution were not significant. Men in the fourth OPG quartile had higher levels of bone resorption markers (11.8-13.1%, P < 0.01-0.001). Conclusions: Men with higher serum OPG concentration had lower cortical thickness and vBMD, probably due to accelerated endo- and intracortical bone turnover, but higher cross-sectional area possibly due to periosteal apposition. Copyright © 2011 by The Endocrine Society.

Schons C.D.,Biomedica | Tavares R.G.,Federal University of Health Sciences, Porto Alegre
Jornal Brasileiro de Patologia e Medicina Laboratorial | Year: 2010

Introduction: The reliability of laboratory results is ensured by the implementation of quality control, which has basic functions, such as analysis, research and prevention of laboratory errors through programs that encompass both internal and external control. Objective: To propose a standard method to use pooled whole blood as internal quality control in the Hematology division. Method: The selected whole blood samples were collected with EDTA, belonged to the same blood group and Rh factor and did not present interfering factors, such as hemolysis, lipemia and icterus. From a total of 30 ml of whole blood it was obtained 3 aliquots of 10 ml each, to which 0 ml (no addition), 1 ml and 5 ml of glycerol (preservative) were added, respectively. The samples were analyzed in the automatic counter ADVIA® 60. After the determination of mean and standard deviation values of all samples, they were evaluated for a period of 45 working days in order to produce a Levey-Jennings graph and to check their stability. Result and conclusion: We could verify that the pool of whole blood prepared in accordance with the methodology proposed does not present the required stability to be used as an alternative internal control in the Hematology division.

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