Analysis for higenamine in urine by means of ultra‐high‐performance liquid chromatography–tandem mass spectrometry: Interpretation of results

Higenamine (Norcoclaurine) is a very popular substance in Chinese medicine and is present in many plants. The substance may be also found in supplements or nutrients, consumption of which may result in violation of anti‐doping rules. Higenamine is prohibited in sport at all times and included in Class S3 (β‐2‐agonists) of the World Anti‐Doping Agency (WADA) 2017 Prohibited List. The presence of higenamine in urine samples at concentrations greater than or equal to 10 ng/mL constitutes an adverse analytical finding (AAF). This work presents a new metabolite of higenamine in urine sample which was identified by means of ultra performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). Samples were prepared according to 2 protocols – a Dilute and Shoot (DaS) approach and a method involving acid hydrolysis and double liquid‐liquid extraction (LLE). To meet the requirements typical for a confirmatory analysis, the screening procedure was further developed. In samples prepared by the DaS method, 2 peaks were observed; the earlier one was specific for higenamine and the later one unknown. MS scan analysis showed mass about 80 Da higher than that of higenamine. In turn, in samples prepared in accordance with the protocol involving hydrolysis, an increase in the area under peak for higenamine was observed, while the second peak was absent. It seems that the described strategy of detection of higenamine in urine avoids false negative results.     

Fenestration and duplication of the vertebral artery

The vertebral artery (VA) acts as a foundation for the posterior circulation of the head and neck. It presents a number of anomalies that can be easily visualized thanks to modern imaging techniques, such as MR and CT angiography or color Doppler ulrasonography. Determining the appropriate terminology for those anomalies can be sometimes more challenging than their recognition. One particular challenge concerns the differentiation betweenVA fenestration and duplication. Because of the different clinical prognoses associated with those anomalies, confusion should be avoided. Knowledge of the morphological anomalies associated with the VA is important for both radiologists and head and neck surgeons, because any injury to the VA can result in a threat to the vascular supply of the brain stem, the cerebral or cerebellar hemispheres, the thalamus, cervical nerve roots and particularly the lateral medulla (Wallenberg's syndrome). This article analyses world literature concerning the issue of VA fenestration and duplication to facilitate a differential diagnosis. Clin. Anat. 26:933–943, 2013. © 2013 Wiley Periodicals, Inc.     

Studies on the effect of lesions of the ventral noradrenergic tract on the antinociceptive action of morphine

Benzazepine (Sch-12679) reduced the shock-induced fighting behavior of rats in a dosedependent fashion. However, the drug also impaired both locomotor behavior and rotorod performance. These results suggest that the aggression suppressive effects of Sch-12679 may be mediated by drug-induced sedation.     

Cardiac mass and function, carotid artery intima media thickness, homocysteine and lipoprotein levels in children and adolescents with growth hormone deficiency

INTRODUCTION: Early atheromatic changes have been found in the carotid and the femoral arteries of young adults with growth hormone (GH) deficiency (GHD). It has been shown that adults with GHD have increased cardiovascular risk factors. The aim of this study was to compare atherosclerotic risk factors in children with GHD and a normal population. PATIENTS AND METHODS: Twenty-five patients (10 girls, 15 boys) with GHD qualified for GH replacement treatment were included in the study. The control group consisted of 22 healthy children (11 girls, 11 boys) with no family history of atherosclerosis and cardiovascular disease. Cardiac mass and function was evaluated by one-dimensional, two-dimensional echocardiography and the Doppler method. Common carotid intima-media thickness was measured with a linear probe (7.5 MHz). Lipid metabolism parameters (total cholesterol, HDL- and LDL-cholesterol, triglycerides), lipoprotein A (Lp(a)) and homocysteine levels were measured. RESULTS: Cardiac dimensions and systolic parameters were within normal values in both groups. The mean index of left ventricular mass (68.37 +/- 18.64 g/m2) in patients with GHD did not differ significantly compared to controls (68.48 +/- 15.56 g/m2). No significant differences between the study and control group were observed when comparing systolic and ejection fraction values. Significantly higher mean values of IVRT parameter were found (70.8 +/- 14.2 ms vs 64.09 +/- 8.54% ms; p < 0.05). Other parameters characterizing systolic function were not significantly different. The mean concentration of total (178.28 +/- 31.1 mg/dl) and LDL-cholesterol (106 +/- 28.68 mg/dl) was significantly higher in patients with GHD than in controls (157.59 +/- 22.39 mg/dl, 84.54 +/- 22.01 mg/dl; p <0.05). Lp(a) (40.34 +/- 12.45 mg/dl vs 11.02 +/- 4.82 mg/dl; p <0.05) and apolipoprotein B (ApoB) (71.12 +/- 18 mg/dl vs 56.72 +/- 11.46 mg/dl; p < 0.05) were significantly higher in patients with GHD. Carotid artery intima-media thickness (IMT) values were significantly higher in patients with GHD compared to healthy children (IMT-L 0.53 +/- 0.058 mm vs 0.41 +/- 0.045 mm; IMT-R 0.54 +/- 0.048 vs 0.42 +/- 0.42 mm, respectively). CONCLUSIONS: No significant differences in parameters of cardiac systolic function and left ventricular mass were found between patients with GHD and healthy children. However, significantly higher Isovolumetric relaxation time (IVRT) values in the group of patients may suggest impaired diastolic function. Significantly elevated IMT values in the common carotid artery were observed in patients with GHD in comparison with healthy children, which may prove the onset of atheromatosis.     

Evaluation of pregnancy-associated plasma protein A (PAPP-A) and free beta subunit of human chorionic gonadotropin (beta hCG) levels and sonographic assesement of fetal nuchal translucency (NT) in singleton pregnancies between 11 and 14 weeks of gestation--Polish multi-centre research

Evaluation of pregnancy-associated plasma protein A (PAPP-A) and free beta subunit of human chorionic gonadotropin (beta hCG) levels and sonographic assessment of fetal nuchal translucency (NT) in singleton pregnancies between 11 and 14 weeks of gestation--Poland's multi-centers research. OBJECTIVES: Pregnancy-associated plasma protein A has been reported to be low in Down syndrome affected pregnancies during the first trimester of pregnancy. Enlarged nuchal translucency (NT) is observed in about 80% of fetuses affected with chromosomal abnormalities and congenital heart defects (CHD). MATERIAL AND METHODS: The aim of this study were to determine value and the medians of free beta-human chorionic gonadotropin (beta-hCG) and pregnancy associated plasma protein-A (PAPP-A) and nuchal translucency thickness in the first trimester in a prospective study of a non-selected Polish population. RESULTS: All examinations have been performed according to the Fetal Medicine Foundation (FMF) rules. We have included 800 women between 11 weeks 0 days and 13 weeks 6 days gestation into a biochemical examination. Women booked into the clinic were offered screening, using a combination of maternal serum free beta-hCG and pregnancy-associated plasma protein-A (PAPP-A) and fetal nuchal translucency thickness. The maternal serum were measured using the Kryptor analyzer (Brahms Diagnostica). All pregnant women have been divided into 2 groups younger than (first group) and older than (second group) 35 years of age. CONCLUSIONS: Nomogrames for free beta-hCG and PAPP-A levels in physiological pregnancy between 11(+0) and 13(6) weeks were determined in the examined population. A positive correlation between PAPP-A and CRL levels, as well as a weak negative correlation between free beta-hCG and CRL, were demonstrated.     

Neural commitment of cord blood stem cells (HUCB-NSC/NP): therapeutic perspectives

Human umbilical cord blood (HUCB) is considered a promising source of neural progenitors capable of being used for cellular therapies in neurological disorders. Here we review briefly our work on the elucidation of mechanisms and development of practical standards as regards the selection, maintenance and use of cord blood derivatives for such purposes. Our results join those of other recent studies in suggesting strongly that, the generation of neural-like cells from tissue belonging to a different germ layer (such as a cord blood is) is most probably explained by reference to a discrete subpopulation of embryonic-like stem cells of pluripotent characteristics. Such cells identified in cord blood through their expression of specific genetic and protein markers can be expanded in vitro and directed toward neurally-committed progenitors differentiating further into more mature neuron-like or macroglia-like cell phenotypes. From this HUCB-derived neural progenitor fraction a novel neural-like stem cell line (HUCB-NSC) has been developed, and characterized in respect of in vitro and in vivo (post-transplantation) properties.     

Expression and localization of Werner syndrome protein is modulated by SIRT1 and PML

Mutations in genes for WRN and BLM RecQ family helicases cause cancer prone syndromes. Werner syndrome, resulting from WRN mutation, is a segmental progeria. Endogenous WRN and BLM proteins localize in nucleoli and in nuclear PML bodies defined by isoforms of the PML protein, which is a key regulator of cellular senescence. We further characterized WRN and BLM localization using labeling with monomeric red fluorescence protein (mRFP). When ectopically expressed, mRFP-WRN (or untagged WRN) forms nuclear bodies, which are donut-shaped in some cells. We identified PML isoforms associating with the nuclear bodies. Interestingly, mRFP-WRN relocalizes from nucleoli to the nucleoplasm, frequently showing conspicuous nucleolar exclusion as well as a decrease in frequency of mRFP-WRN nuclear bodies in response to overexpression of wild-type and deacetylase mutant (H363Y) SIRT1 proteins. Similar nucleolar relocalization in response to wild-type SIRT1 was detected for mRFP-labeled BLM. Moreover, increased SIRT1 expression was associated with the downregulation of endogenous WRN and a decreased frequency of cells with BRCA1 foci. Our data indicate for the first time that SIRT1 protein may be functionally associated with WRN and BLM helicases and that some major SIRT1 functions may not require its deacetylase activity.