Enrichment techniques and trace analysis with microbore columns in liquid chromatography

The advantages of microbore columns for trace analysis by liquid chromatography are identified, with reference to on-column enrichment techniques performed on analytical micro-columns. The selectivity and high sensitivity of the amperometric detector are utilized in combination with a microbore column for a number of pharmaceutical and bioanalytical analyses, including phenothiazines, parabens, sulphonamides, catecholamines, tetracyclines, vitamins, amino acids and dipeptides.     

Upregulation of asparagine synthetase fails to avert cell cycle arrest induced by L-asparaginase in TEL/AML1-positive leukaemic cells.

L-Asparaginase is a standard component in chemotherapy of childhood acute lymphoblastic leukaemia (ALL). Leukaemic cells carrying TEL/AML1 fusion gene are more sensitive to treatment with L-asparaginase compared to other subtypes of ALL. We demonstrate in vitro the prolonged growth suppression of TEL/AML1[+] cells compared to TEL/AML1[-] leukaemic cells after L-asparaginase treatment simulating treatment protocol. Cell cycle analysis revealed TEL/AML1[+] cells to accumulate in G1/G0 phase (81-98%) compared to TEL/AML1[-] cells (47-60%). Quantitative analysis of asparagine synthetase (AsnS) expression showed the ability of TEL/AML1[+] cells to increase AsnS mRNA levels after L-asparaginase treatment to the same extent as TEL/AML1[-] leukaemic and nonleukaemic lymphoid cells. We hypothesise that TEL/AML1[+] cells are unable to progress into the S phase of cell cycle under nutrition stress caused by L-asparaginase, despite the ability of AsnS upregulation. Significantly higher expression of AsnS was found in untreated leukaemic cells from children with TEL/AML1[+] ALL (n=20) in comparison with the group of age-matched children with ALL bearing no known fusion gene (n=25; P=0.0043). Interestingly, none of the TEL/AML1[+] patients with high AsnS level relapsed, whereas 10/15 patients with AsnS below median relapsed (P=0.00028). Therefore, high AsnS levels in TEL/AML1[+] patients correlate with better prognosis, possibly reflecting the stretched metabolic demand of the lymphoblast.     

Fluorescence intensities of composite resins on photo images

Odontology - Recording fluorescence using flash photography, may help reduce time of capture and apply effectively in clinical practice. To test methods for visualizing composite resins...     

Kinetics of bilirubin and liver enzymes is useful for predicting of liver graft-versus-host disease

Graft-versus-host disease (GVHD) is the most frequent complication after allogeneic hematopoietic cell transplantation. We analyzed the kinetics of bilirubin and liver enzymes in 47 cases with liver GVHD and in 47 cases without GVHD after allogeneic transplantation for various hematological malignancies. The duration of an liver GVHD episode (LGVHD) was defined as the interval from the point when the criteria of LGVHD were met to the decrease to < 2 upper normal limit (UNL) for aminotransferases or bilirubin < 34 μmol/l for bilirubin. The imminent LGVHD episode was defined as the interval from the start of continuous increase (≥ 3 consecutive rising values) of bilirubin and liver enzymes above UNL to the point of LGVHD diagnosis.The number of imminent LGVHD episodes, and median length in days were as follows: bilirubin (39;5), ALT(28;12), AST(9;12), GGTP(34;9), and ALP(13;14). Statisticallly significant associations between asymptomatic continuous increase of bilirubin, ALT, and GGTP and later liver GVHD manifestation were found (p=0.004, p=0.008, p=0.005, respectively). The asymptomatic continuous increase in bilirubin, ALT, and GGTP occurred at a median of 5, 12, and 9 days before liver GVHD episode, respectively. In the control group without GVHD, median levels of bilirubin and liver enzymes were within normal limits and no continuous increase was observed.Kinetics of bilirubin and liver enzymes is useful for predicting of liver GVHD. A continuous increase of bilirubin and/or ALT, GGTP before the standard liver GVHD criteria are met can be a sign of coming liver GVHD.     

The competition between enamel and dentin adhesion within a cavity: An in vitro evaluation of class V restorations

To gain more insight into the consequences of curing contraction within the tooth cavity, we assessed the margin behavior of 12 contemporary restorative systems in class V restorations with margins located on enamel and dentin after mechanical loading and water storage. Mixed class V cavities were prepared on extracted human molars and restored using five etch and rinse and seven self-etch adhesive systems with their corresponding composites. Marginal adaptation was evaluated by using a computer-assisted quantitative marginal analysis in a scanning electron microscope (SEM) on epoxy replicas before, after thermal and mechanical stressing and after 1 year of water storage. The interactions of "testing conditions", "adhesive-composite combination" and "tooth substrate" with "marginal adaptation" were evaluated by two-way ANOVA. Fatigue, stress and storage conditions had significant effects on the marginal adaptation. Only two groups (Optibond FL and G Bond) presented equal percentages of marginal adaptation on enamel and dentin; in the other groups, the rate of degradation was product dependent. All materials tested showed a distinct behavior on enamel and dentin. In addition to mechanical resistance and long-term stability, differences within materials also exist in their ability to simultaneously bond to enamel and dentin.     

Postsynaptic muscarinic m2 receptors at cholinergic and glutamatergic synapses of mouse brainstem motoneurons

In many brain areas, few cholinergic synapses are identified. Acetylcholine is released into the extracellular space and acts through diffuse transmission. Motoneurons, however, are contacted by numerous cholinergic terminals, indicating synaptic cholinergic transmission on them. The muscarinic m2 receptor is the major acetylcholine receptor subtype of motoneurons; therefore, we analyzed the localization of the m2 receptor in correlation with synapses by electron microscopic immunohistochemistry in the mouse trigeminal, facial, and hypoglossal motor nuclei. In all nuclei, m2 receptors were localized at the membrane of motoneuronal perikarya and dendrites. The m2 receptors were concentrated at cholinergic synapses located on the perikarya and most proximal dendrites. However, m2 receptors at cholinergic synapses represented only a minority (<10%) of surface m2 receptors. The m2 receptors were also enriched at glutamatergic synapses in both motoneuronal perikarya and dendrites. A relatively large proportion (20–30%) of plasma membrane–associated m2 receptors were located at glutamatergic synapses. In conclusion, the effect of acetylcholine on motoneuron populations might be mediated through a synaptic as well as diffuse type of transmission. J. Comp. Neurol. 521:2008–2024, 2013. © 2012 Wiley Periodicals, Inc.     

Minimal residual disease prior to stem cell transplant for childhood acute lymphoblastic leukaemia

Allogeneic stem cell transplantation (SCT) is a highly effective therapy for childhood acute lymphoblastic leukaemia (ALL). Concerns about unnecessary toxicity and expense mean that SCT is currently largely reserved for children who cannot be cured with chemotherapy. Not surprisingly, many such children also fail SCT. Retrospective studies have shown that a single analysis of minimal residual disease (MRD) pre-SCT identified those at highest risk of relapse. It is now appropriate to call for the universal incorporation of standardized MRD testing into SCT protocols as the next step to maximize the clinical impact of this technology in ALL.