Biomedical subjects
Publications and source records attributed to B Xie.
Determination of chloramphenicol in the ear drops by first derivative spectrometry was studied. The detecting wavelength was 303.0 nm. The linear range was 0-65.2 micrograms.ml-1 (r = 0.9999). The average recovery was 99.84%, RSD = 0.37% (n = 8). The method is simple and accurate.
"Spongy left ventricular myocardium," or noncompaction of left ventricular myocardium, is a rare disorder of endomyocardial morphogenesis. It is usually seen in the pediatric population and is often associated with other congenital cardiac malformations. We describe an adult with noncompaction of left ventricular myocardium without associated congenital cardiac anomalies.
Regulatory proteins are often ubiquitinated, depending on their phosphorylation status as well as on their association with ancillary proteins that serve as adapters of the ubiquitination machinery. We previously demonstrated that c-Jun is targeted for ubiquitination by its association with inactive c-Jun NH2-terminal kinase (JNK). Phosphorylation by activated JNK protects c-Jun from ubiquitination, thus by prolonging its half-life. In the study reported here, we determined the ability of JNK to target ubiquitination of its other substrates (Elk1 and activating transcription factor 2 (ATF2)) and associated proteins (ATF2 and JunB). We demonstrate that phosphorylation by JNK protects ATF2, but not Elk1, from JNK-targeted ubiquitination. We also show that association of inactive JNK with JunB or ATF2 is necessary to target them for ubiquitination. Unlike its targeting of c-Jun, JNK requires additional cellular components, yet to be identified, to target the ubiquitination of ATF2. Elk1 is phosphorylated by JNK, but JNK neither associates with nor targets Elk1 for ubiquitination. The implications for the dual role of JNK in the regulation of ubiquitination and stability of c-Jun, ATF2, and JunB in normally growing versus stressed cells are discussed.
A miniaturized thermal biosensor has been evaluated as part of a flow-injection analysis system for the determination of glucose in whole blood. Glucose was determined by measuring the heat evolved when samples containing glucose passed through a small column with immobilized glucose oxidase and catalase. Samples of whole blood (1 microliter) can be measured directly, without any pretreatment. The correlation in the response between the thermal biosensor, the Reflolux S meter (Boehringer Mannheim), the Granutest 100 glucose test kit (Merck Diagnostica) and the Ektachem (Kodak) instrument was evaluated. The influence of the hematocrit value and of possible interferences is reported. The correlation measurements show that the thermal biosensor calibrated with aqueous glucose standards generally gives lower values on blood glucose than the reference methods calibrated for serum or blood measurements. Mean negative biases range from 0.53 to 1.16 mmol/l. Differences in sample treatment clearly complicate comparisons and the proper choice of reference method. There was no influence from substances such as ascorbic acid (0.11 mmol/l), uric acid (0.48 mmol/l), urea (4.3 mmol/l) and acetaminophen (0.17 mmol/l) on the response to 5 mmol/l glucose. The hematocrit value does not influence the glucose determination, for hematocrit values of between 13 and 53%.
The G(M2) activator protein is a substrate specific cofactor for degradation of G(M2) ganglioside by lysosomal beta-hexosaminidase A. Mutations in the gene encoding the activator result in the AB-variant form of G(M2) gangliosidosis. The activator protein contains at least three functional elements; a hydrophobic binding pocket, an oligosaccharide binding site(s), and an area that interacts with hexosaminidase A. In this report a fluorescence dequenching assay specific for only the hydrophobic binding pocket is evaluated and optimized. It is shown that various glycolipids inhibit the transport between liposomes of a self-quenching fluorescent lipid probe, octadecylrhodamine, by the activator protein. The level of inhibition produced by each glycolipid is then used to characterize the oligosaccharide-binding specificity of the activator. The fluorescence dequenching assay is also used to evaluate the functionality of a truncated form of the activator protein. Our results indicate that this simple assay can be used to determine structure-function relationships within the normal or mutant forms of the activator. The data suggest that the C-terminus of the activator is required to produce a functional hydrophobic binding pocket.
The achondroplasia class of chondrodysplasias comprises the most common genetic forms of dwarfism in humans and includes achondroplasia, hypochondroplasia and thanatophoric dysplasia types I and II (TDI and TDII), which are caused by different mutations in a fibroblast growth-factor receptor FGFR3 (ref. 1). The molecular mechanism and the mediators of these FGFR3-related growth abnormalities are not known. Here we show that mutant TDII FGFR3 has a constitutive tyrosine kinase activity which can specifically activate the transcription factor Stat1 (for signal transducer and activator of transcription). Furthermore, expression of TDII FGFR3 induced nuclear translocation of Stat1, expression of the cell-cycle inhibitor p21(WAF1/CIP1), and growth arrest of the cell. Thus, TDII FGFR3 may use Stat1 as a mediator of growth retardation in bone development. Consistent with this, Stat1 activation and increased p21(WAF1/CIP1) expression was found in the cartilage cells from the TDII fetus, but not in those from the normal fetus. Thus, abnormal STAT activation and p21(WAF1/CIP1) expression by the TDII mutant receptor may be responsible for this FGFR3-related bone disease.
In this review a universal thermistor-based biosensor system is described with examples from clinical chemistry, bioprocess monitoring and environmental control. The technique is based on the measurement of the small temperature changes associated with enzymatic reactions occurring in a microreactor with immobilized enzyme. The system has good operational stability and a sensitivity that permits measurements down to 1 microM concentrations. Current developments include devices constructed by micromachining for multisensing purposes and miniaturised instrumentation intended for use in portable monitoring. With use of special supports for enzyme immobilisation even untreated whole blood samples can be applied. Another current line of investigation involves hybrid biosensors, such as combinations of electrochemistry and calorimetry into bioelectrocalorimetric devices with interesting new properties.
Bovine leukaemia virus (BLV) proviral DNA was detected in lymphocytes isolated from cattle with persistent lymphocytosis (PL) by the polymerase chain reaction (PCR) and in-situ hybridization (ISH) with a biotinylated pX DNA probe. Many positive cells were observed when short-term culture and a combination of ISH with PCR were used. Immunohistochemical examination of lymphocytes isolated from the lymph node showed that BLV attached mainly to surface immunoglobulins (SIg) of positive B lymphocytes, and to a few tumour-associated antigen (TAA)-, PanT-, and CD8-positive cells and non-CD4 positive cells. Electron microscopical examination revealed colloidal gold particles within the nuclei and cytoplasm of lymphocytes. Lymphoid cells from neoplastic lymph node of enzootic bovine leukosis (EBL) cases gave particularly strong positive signals with the ISH-PCR method. The technique of combined ISH and PCR with a biotinylated pX probe may prove useful in future studies of EBL.
Milrinone is an inotropic drug with vasodilator activity that has been shown to be useful in increasing cardiac output and decreasing wedge pressure. Despite these advantages, it is unknown whether this drug can be used for the treatment of perioperative spasm of coronary bypass grafts. This study was undertaken to investigate the in vitro vascular effect of milrinone on internal thoracic arteries obtained from patients undergoing coronary artery bypass grafting. The results showed that milrinone produced a potent, concentration-dependent, preventive effect on the norepinephrine-induced contraction of internal thoracic arteries, as well as reversing contraction of internal thoracic arteries by receptor-dependent agents, including the thromboxane A2 mimetic U46619, the vasoconstrictor peptide endothelin-1, and the alpha1-adrenal receptor agonist phenylephrine. The relaxing effect of milrinone was weaker, however, on internal thoracic arteries contracted with 25 mmol/L potassium chloride. Comparison of milrinone with other vasodilators, including papaverine, nitroprusside, and glyceryl trinitrate, showed milrinone to be more potent than papaverine but less potent than nitroprusside and glyceryl trinitrate. The inhibitory effect of milrinone on internal thoracic artery contraction appeared as a reduction in contractile force, not as an increase in the values of concentrations of the agonists causing 50% maximal contraction, which indicates that milrinone exerts its vasodilator effect directly on the smooth muscles, not on the membrane receptors. The results also showed no significant difference in relaxing effect between internal thoracic artery rings with and without endothelium. In conclusion, this study provides experimental evidence that milrinone is a potent, endothelium-independent, direct vasodilator of the human internal thoracic artery and provides the scientific rationale for a future clinical trial with this drug for the perioperative treatment of internal thoracic artery spasm in cardiac surgical patients.
Around 20% of patients with third generation implantable cardioverter defibrillators receive inappropriate therapy, usually triggered by atrial fibrillation. This is because the criteria used for ventricular tachycardia detection by current implantable cardioverter defibrillators are based on the analysis of a sequence of RR intervals and may be inappropriately satisfied by supraventricular tachyarrhythmias. Algorithms for ventricular tachycardia detection were challenged against the full RR interval sequences from 482 spontaneous episodes of atrial fibrillation and 260 spontaneous episodes of ventricular tachycardia to determine their ability to discriminate between the arrhythmias. The sensitivities and specificities of the algorithms were calculated over a wide range of programmable parameters. For a given window length and detection interval, the most stringent algorithms, that required all beats to be classified as "fast", were more specific than those allowing a proportion of "normal" intervals, even after adjustment for differing sensitivity. These differences were less marked for faster tachycardias. Specificity increased with the detection window length to a limit of approximately 18 beats. We conclude that ventricular tachycardia is detected with the highest specificity if all beats in an analyzed sequence are required to be "fast," even after lengthening of the tachycardia detection interval to maintain sensitivity. Further improvement in algorithm performance may require the incorporation of criteria such as tachycardia onset and stability.
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OBJECTIVE: To identify possible factors associated with primary failure of radiofrequency ablation of accessory pathways or recurrence of accessory pathway conduction. PATIENTS AND METHODS: Radiofrequency ablation of accessory pathways failed in 25 of 243 patients, and recurrence of accessory pathway conduction occurred in an additional 13 patients. Factors possibly related to primary failure and recurrence were analysed. RESULTS: Primary failure and recurrence were less frequent in patients with left sided pathways (7% v 19%; 4% v 24%; P = 0.04). The factors that might relate to primary failure included an unstable catheter position (seven patients), a possible epicardial pathway (six patients), or misdiagnosis of accessory pathway location (two patients). The major factors for recurrence included the stability of the local atrial electrogram < or = 0.5 together with the stability of the local ventricular electrogram < or = 0.8, and prolonged time to pathway conduction block > or = 12 seconds). Thirty one patients underwent repeat ablation which was successful in 28. CONCLUSIONS: Primary failure and recurrence were more frequent in patients with right sided pathways. An unstable catheter position and a possible epicardial pathway location are the main contributing factors for primary failure, while unstable local electrograms and prolonged time to block are independent predictors for recurrence.
The mammary tissues of 6 cows with bovine leukemia virus (BLV) antibody and subclinical mastitis were investigated histopathologically, and their organ cultures were ultrastructurally observed. Numerous BLV particles, 110 to 120 nm in diameter, were seen around lymphocytes, which had infiltrated into mammary alveoli and showed blastogenesis under culture. Particles budding from the cell membrane were also found.
A survey on nutritional knowledge, attitude and practice (K-A-P) was carried out among 965 residents aged between 20 and 50 and randomly selected in Beijing, Guangzhou and Shanghai. The results showed that subjects' knowledge level was not high, and their knowledge scores were significantly correlated with their education levels (r = 0.3011, P < 0.010). Subjects aged above 35 had a higher knowledge level than that of younger ones. Subjects from three cities held a positive and favorable attitude towards the acquisition of nutritional knowledge, reasonable dietary pattern and healthy diet habits. Results of food frequency survey indicated that the consumption of rice/flour, meat/poultry, vegetables, fruits, milk and milk products was relatively high. In addition, 89.5%, 78.8% and 47.8% of subjects selected newspaper, TV and broadcast as their main sources of nutritional information respectively. This survey provided basis on content and method for future nutrition education.
To find out a method to predict lower body negative pressure (LBNP) tolerance with noninvasive physiological measurements, 11 healthy young subjects were tested in an LBNP chamber in sitting position. Incremental negative pressure was used for evaluating LBNP tolerance time. End points of the test were onset of presyncopal symptoms or completion of 15 min test. According to their reactions, the 11 subjects were divided into good tolerance group (group A, n = 5) and poor tolerance group (group B, n = 6); At the 11th minute of LBNP, mean arterial pressure (MAP), magnitude of decrease of regional cerebral oxygen saturation (delta SrO2) and cardiac output (delta CO) showed significant difference between group A and B(P <0.05); Mean error was 0.54 +/- 0.42 min when LBNP tolerance was predicted using MAP and delta SrO2 at this moment by regression analysis.
The purpose of this study was to observe and analyse the changes of regional cerebral oxygen saturation (SrO2) under lower body negative pressure (LBNP). 12 healthy young subjects were tested in an LBNP chamber in sitting position. Incremental negative pressure was used and end points of test were onset of presyncopal symptoms or completion of the 15min test. The results were: (1) SrO2 showed a significant decrease under LBNP; (2) The magnitude of decrease of SaO2 showed significant differences among subjects with different reactions at the termination of LBNP; (3) Under LBNP the arterial oxygen saturation (SrO2) kept constant. The changes of SrO2 correlated strongly with the percentage changes of superaorbitalis arterial blood flow (r = 0.59-0.86, P < 0.05 or 0.01). It was suggested that the changes of SrO2 were related to the decrease of cerebral blood flow under LBNP. It was possible to use SrO2 to detect presyncopal and syncopal symptoms. So the monitoring of SrO2 could be used in studies of G-LOC.
The volatile compounds of three cultivars of hawthorn were studied. Changko (Crataegus pinnatifida Bge.) hawthorn fruit was harvested after ripening from Hubei Province and those of Heihong and Dajinxing were from Shandong Province. The volatile compounds of each hawthorn cultivars were obtained by SDE (simultaneous distillation-extraction) equipment, by using CH2Cl2 as extracting solvent. The volatile extract was concentrated at 40-50 degrees C under vacuum to 0. 05mL or so and was ready for GC and GC/MS analysis. A DB-Wax fused silica capillary column (50m x 0.32mm i.d.; 1microm thickness) and a flame ionization detector (FID) was employed in GC analysis. The temperature program included of a 5 min isothermal period at 40 degrees C, temperature increases of 2 degrees C/min from 40 degrees C to 240 degrees C, and a 60 min isothermal period at 240 degrees C. Mass spectra were obtained by electron impact at 70eV and a source temperature of 250 degrees C. Thirty-two volatile compounds of the hawthorn fruit were identified, which comprised 61%-68% of the volatile fraction. The ten major components were cis-3-hexenol, cis-3-hexenyl acetate, alpha-terpineol, furfural, hexanol, hexyl acetate, nonanal, citral, 3-penten-2-one and trans-2-decenal. The molecular weight range of the major volatile fraction covers from C3 to C10. Both qualitative and quantitative differences in the volatile constituents among the three cultivars were not remarkable.
The structure-function study of hydroxysteoid dehydrogenases has stimulated the development of their chromatography, which in turn reveals more mechanisms of these enzymes. Due to the various membrane associations and mild hydrophobic nature of most of the enzymes studied up to now, hydrophobic interaction chromatography has played a crucial role in their purification, using media such as phenyl-Superose or Sepharose-PEG. At the same time, affinity chromatography, especially the dye-containing columns, proves very efficient for these dehydrogenases, as the latter utilizes adenylyl-containing cofactors. Elution by their specific ligand facilitates their purification. In this paper, the use of detergents in the purification of these enzymes is also reviewed. Hydroxysteroid dehydrogenase preparation is further improved by rapid purification which facilitates the elimination of protein microheterogeneity, caused in vitro by oxidation, reduction or partial proteolysis. This process was shown to increase the crystallizability of the enzymes [Lin et al., J. Cryst. Growth, 122 (1992) 242-245; Zhu et al., J. Mol. Biol., 234 (1993) 242-244]. The fast purification permitted a simpler procedure and better combination of various columns than conventional chromatography. This leads to even higher efficiency, yielding homogeneous and highly active preparations.