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Biomedical subjects

C Lu

Publications and source records attributed to C Lu.

At least 19 recordsLinked to original sources

Characterization of small nontranslated polyadenylylated RNAs in vaccinia virus-infected cells.

Host protein synthesis is selectively inhibited in vaccinia virus-infected cells. This inhibition has been associated with the production of a group of small, nontranslated, polyadenylylated RNAs (POLADS) produced during the early part of virus infection. The inhibitory function of POLADS is associated with the poly(A) tail of these small RNAs. To determine the origin of the 5'-ends of POLADS, reverse transcription was performed with POLADS isolated from VV-infected cells at 1 hr and 3.5 hr post infection. The cDNAs of these POLADS were cloned into plasmids (pBS or pBluescript II KS +/-), and their nucleotide composition was determined by DNA sequencing. The results of this investigation show the following: There is no specific gene encoding for POLADS. The 5' ends of POLADS may be derived from either viral or cellular RNAs. Any RNA sequence including tRNAs, small nuclear RNAs and 5'ends of mRNAs can become POLADS if they acquire a poly(A) tail at their 3' ends during infection. This nonspecific polyadenylylation found in vaccinia virus-infected cells is probably conducted by vaccinia virus poly(A)+ polymerase. No consensus sequence is found on the 5' ends of POLADS for polyadenylylation. The 5' ends of POLADS have no direct role in their inhibitory activity of protein synthesis.

Cell-Free System

Synthesis of optically pure chrysobactin and immunoassay development.

Chrysobactin (alpha-N-(2,3-dihydroxybenzoyl)-D-lysyl-L-serine), a siderophore that is essential for systemic virulence by plant pathogenic Erwinia chrysanthemi, was synthesized with high diastereomeric purity. Chrysobactin was prepared by coupling the N-hydroxysuccinimide ester of alpha-N-(2,3-dibenzyloxybenzoyl)-epsilon-N-Cbz-D-lysine with L-serine benzyl ester followed by deprotection via hydrogenolysis. Optically pure chrysobactin was obtained with 98% overall yield. A monoclonal antibody to ferric chrysobactin was developed and characterized as IgM. The antibody reacts with chrysobactin, ferric chrysobactin and less strongly with ferric dihydroxybenzoic acid. The antibody reacts weakly with the siderophores ferrichrome, A, ferric pseudobactin and ferric rhodotorulic acid. This antibody was used in a competitive immunoassay to detect ferric chrysobactin at 10(-8) to 10(-10) mol. This immunoassay may provide a useful method for the detection of chrysobactin in plant samples.

Animals

Duodenum as a immediate brake to gastric outflow: a videofluoroscopic and manometric assessment.

BACKGROUND & AIMS: Duodenal infusion of HCl or lipid delays gastric emptying. The aim of this study was to assess whether this delay was in part caused by mechanical activity of the duodenum. METHODS: Synchronized videofluoroscopy and manometry was used in 8 volunteers (5 men and 3 women) to examine contractile and flow patterns during duodenal infusion of 0.9% NaCl, HCl, 5% NaCl, bile, and sodium oleate, each mixed with 20% (wt/vol) barium sulfate. RESULTS: Within 15-30 seconds of infusion, HCl and 5% NaCl induced frequent large-amplitude contractions greater tha those induced by 0.9% NaCl. Initially, there was rapid dispersion of HCl followed by prolonged, tonic occlusion of the duodenum. The duodenal diameter decreased compared with that observed during 0.9% NaCl or oleate infusion. In contrast, after infusion of oleate or bile, duodenal diameter increased and there were fewer, smaller-amplitude, nonpropagating contractions with prolonged retention of solutions. Barium (20%; wt/vol) did not influence the motility index of any solution. CONCLUSIONS: HCl and 5% NaCl may restrict gastric outflow by inducing tonic occlusion of the duodenum, whereas bile and lipid may delay clearance by decreasing duodenal tone and contractility. Thus, the duodenum may serve as an immediate brake to gastric outflow either by delaying clearance or by offering rapid tonic resistance.

Adolescent

Manometric responses of human duodenum during infusion of HCl, hyperosmolar saline, bile and oleic acid.

Duodenal motor activity is incompletely understood. The purpose of this study was to define the contractile patterns of the duodenum that occur in response to rate controlled injection of various solutions. In nine healthy volunteers we placed a six channel perfused catheter, and recorded pressure activity in the antrum, pylorus and duodenum. Volumes of 10 and 20 mL of 0.9% NaCl, 100 mM HCl (pH 1), 5% NaCl (1711 mOsm/kg), human bile and iso-osmolar sodium oleate were randomly injected into the duodenum at 20 ml/min, starting 15 min after phase III migratory motor complex (MMC). A 20 mL bolus of each solution caused more activity (P < 0.05) than a 10 mL bolus, but the motor pattern was similar. The control, 0.9% NaCl, produced occasional pressure waves, whereas bile and sodium oleate induced more (P < 0.05) activity which consisted of low amplitude, isolated or clusters (2-4 cycle/min) of non-propagating pressure waves that occurred at random sites. In three subjects, oleate produced isolated pyloric phasic contractions. In contrast, HCl and 5% NaCl induced high amplitude pressure waves that were seen either at a single channel or at multiple channels, occurring simultaneously. The motility index was also greater (P < 0.05) than that induced by other solutions. Additionally, within 2 min of infusion, a phase III MMC like pattern was observed in five of the nine subjects who received HCl and three of the nine who received 5% NaCl. A non-nutrient iso-osmolar solution induced occasional motor activity. HCl and hyperosmolar solutions induced more frequent and large amplitude, segmental contractions whereas lipid and bile induced fewer and smaller amplitude contractions. The volume, the pH, the osmolar and the nutrient make up of the infusate may each influence the duodenal motor responses.

Adolescent

Volume shifts, unfolding and rolling of haustra in the isolated guinea pig caecum.

The large intestine extracts water from chyme and compacts chyme into faecal conglomerates; it is unclear what role the special pockets known as colonic haustra have in these events. Here we monitored the movements of haustra in isolated preparations of guinea pig caecum using videocamera and ultrasound and related them to contractions of muscle flaps and movements of glass beads in haustral pockets. We found that in partially filled caecal loops localized contractions of taeniae shift volume back and forth between adjacent haustra; volume unfolds haustral walls in a characteristic sweep with sequential intrahaustral folds popping out; cyclic contractions and relaxations of the fold then produce the caterpillar-like movement known as haustral rolling; ultrasound showed that haustral rolling made the haustral flow channel narrower and longer as haustral folds increase their height from 7.5 +/- 1.5 mm to 16 +/- 4 mm and their distance from 4.1 +/- 0.2 mm to 7.9 +/- 0.3 mm; luminal contents were alternatively shaken off the haustral wall, whirled around the lumen or left to settle. We also suspended the row of haustra between two taeniae inside a frame and attached flaps of taeniae and haustral folds to strain gauges to record their mechanical activity; both taeniae and haustral folds produced an undulating baseline tension; during rolling, folds produced phasic contractions at 17 +/- 2 cycles min-1 which propagated distally across haustral septa; rolling constantly shuffled around glass beads placed inside the haustra. When we stimulated the intramural nerves to the caecum through bipolar electrodes, all contractile activity was temporarily inhibited and haustral septa flattened; a rebound contraction then propagated aborally from the caecal pole and swept the glass beads ahead of it. Thus, tonic contractions of taeniae shift caecal contents back and forth across haustral septa; expansion of haustra triggers haustral rolling which shuffles contents; both these movements produce local flow within and between haustra which might enhance the separation of solid and liquid colonic contents.

Animals

Image reconstruction with acoustic measurement using distorted Born iteration method.

The distorted Born iterative method (DBIM) is applied to solve electromagnetics and ultrasonics inverse scattering problems. First, we use the DBIM to process the data, which are the scattered fields from two-dimensional cylinders with arbitrary cross section. From this simulation, we confirmed that the first-order Born approximation can be applied to larger objects as long as the phase change of a wave passing through the object due to its presence is smaller than a limit. Then we applied DBIM to process the ultrasonic measurement data. Images for a balloon and an egg that are immersed in water have been reconstructed and compared with those from the first-order diffraction tomography (DT).

Humans

[A PCR marker-based selection for resistance to bacterial blight in rice].

Molecular marker-based selection in plant breeding requires not only suitable molecular markers closely linked to the known genes, but a simple, economic and reliable analyzing technique. We report here a useful PCR marker for genetic diagnostics in breeding for resistance to rice bacterial blight. Xa21 is a newly found gene of rice which confers resistance to bacterial blight caused by Xanthomonas oryzae pv. oryzae. We produced two F2 populations between one resistant line IRBB21 containing Xa21 and two susceptible varieties, respectively. One PCR marker, PB78, detected polymorphism between the susceptible varieties and the resistant line. Cosegregation between Xa21 and PB78 was studied in the two F2 populations. The results showed that Xa21 was closely linked to the molecular marker, the crossing over value was 2.48%. Marker-based selection revealed that 100% of the plants with homozygous resistant genotype of PB78 showed resistance to bacterial blight. The available approaches detecting molecular markers in plant breeding are also discussed.

Base Sequence

Dipyridamole echocardiography for diagnosis of coexistent coronary artery disease in hypertrophic cardiomyopathy. Echo-Persantine International Cooperative (EPIC) Study Group--Subproject Hypertrophic Cardiomyopathy.

The recognition of coexistent coronary artery disease (CAD) in patients with hypertrophic cardiomyopathy may be difficult by noninvasive testing based upon electrocardiographic changes or perfusion defects. Dipyridamole-stress echocardiography has proved a sensitive and highly specific test for noninvasive diagnosis of CAD in various patient subsets. To establish the feasibility, safety, and diagnostic accuracy of dipyridamole-stress echocardiography in patients with hypertrophic cardiomyopathy, we performed high-dose dipyridamole testing (up to 0.84 mg/kg over 10 minutes) in 88 patients with hypertrophic cardiomyopathy (63 men; mean age +/- SD, 46 +/- 17 years). A subset of 60 patients was referred for coronary angiography independently of test results; CAD was defined as > or = 50% diameter narrowing in at least 1 major coronary vessel. Dipyridamole echocardiography/electrocardiography testing was completed in all patients, with no limiting side effects or adverse reactions. In the subgroup of 60 patients with coronary angiography (14 with and 46 without CAD), chest pain occurred in 18 patients (8 with and 10 without CAD, p = NS); ST-segment depression > or = 2 mm from baseline in 28 (7 with and 21 without CAD, p = NS); and transient dyssynergy in 10 patients (10 with and none without CAD, p < 0.0001). Assuming the transient regional dyssynergy to be the only criterion of positivity, the dipyridamole echocardiography test showed 71% sensitivity, 100% specificity, 100% positive predictive value, and 93% diagnostic accuracy for diagnosis of angiographically assessed CAD. We conclude that high-dose dipyridamole echocardiography testing may be considered a feasible and accurate tool for the noninvasive diagnosis of CAD in patients with hypertrophic cardiomyopathy.

Adult

Complex coronary artery lesion morphology influences results of stress echocardiography.

BACKGROUND: The likelihood of a positive response with dipyridamole stress echocardiography (DSE) is directly related to the extent and severity of angiographically assessed coronary artery disease. Whether coronary lesion morphology--a known predictor of adverse cardiac events--may also modulate stress echo results remains unknown. The objective of our study was to assess the relation between stenosis lesion morphology and stress echocardiographic results. METHODS AND RESULTS: High-dose (up to 0.84 mg/kg over 10 minutes) DSE and coronary angiographic data of 68 in-hospital patients (39 with stable angina, 29 with angina at rest) with nonoccluding, single-vessel disease at angiography and no previous myocardial infarction were analyzed. DSE was performed in all patients within 3 days of coronary angiography. An angiographic lesion was considered complex when irregular borders and/or intraluminal lucencies suggestive of ulcer and/or thrombus were present. According to angiographic lesion morphology, two groups were identified: group 1, with simple coronary lesions, and group 2, with complex coronary lesions. The two groups were matched for number of patients (n = 34 in each group), age (group 1, 59 +/- 9 versus group 2, 59 +/- 10 years, P = NS), and coronary artery stenosis severity by quantitative coronary angiography (group 1, 60 +/- 7% versus group 2, 58 +/- 6% diameter reduction, P = NS). The sensitivity of DSE was lower in patients of group 1 when compared with group 2 (53% versus 85%, P < .001). Among positive DSE, the low-dose (0.56 mg/kg over 4 minutes) positivity was less frequent in group 1 than in group 2 patients (17% versus 62%, P < .01). Exercise ECG was completed in 66 patients, and it was positive (> .1 mV ST-segment shift from baseline) in 20 out of 33 group 1 and in 22 out of 33 group 2 patients (61% versus 67%, P = NS). The peak rate-pressure product tended to be higher in group 1 than in group 2 patients (257 +/- 52 versus 240 +/- 64 mm Hg x beats per minute x 10(2), P = NS). CONCLUSIONS: In patients with single-vessel disease without coronary occlusion or previous myocardial infarction, coronary lesion morphology of the complex type is associated with a higher DSE sensitivity and with a greater prevalence of low-dose, positive responses. Presence of irregular plaque contours, not only plaque geometry, is important in modulating stress responses in the presence of angiographically assessed coronary artery disease.

Aged

Determination of mutational spectrum of the pesticide, captan, with an improved set of Escherichia coli LacZ mutants.

The mutational spectrum of the fungicide, captan, was determined using a set of improved Escherichia coli lacZ mutants. Captan created mutations mostly at dA-dT sites (83%) with only 17% occurring at dG-dC sites. The hydrolysis products of captan do not appear to be mutagenic because samples of captan at different hydrolysis stages showed basically the same mutational spectra: 31% at AT --> CG transversions, 8% of GC --> AT transitions, 2% of GC --> CG transversions, 8% of GC --> TA transversions, 19% of AT --> TA transversions, and 32% of AT --> GC transitions. Prepared solutions of captan lost their mutational activity gradually over time, indicating that the rate of decrease in mutagenicity agreed with the kinetics of captan hydrolysis reported in other studies. Using the change in mutagenicity to predict degradation, the hydrolysis of captan in pH 7.0 buffer was about three times faster than the hydrolysis carried out in pH 4.5 buffer. To our knowledge, this is the first presentation of mutational spectrum of captan.

Base Sequence

Preparation and a structure-function analysis of human ciliary neurotrophic factor.

Ciliary neurotrophic factor (CNTF) is a trophic protein that promotes survival and/or differentiation of a variety of neuronal cell types including sensory, sympathetic, and motor neurons. CNTF, leukemia inhibitory factor (LIF), interleukin-6 (IL-6) and oncostatin M (OSM) share a predicted common helical framework and partially identical receptor components. In this study, we present the preparation and structure--functional analysis of recombinant human CNTF. The human CNTF gene was expressed under the control of the PL promoter in Escherichia coli, and the mutants were constructed by insertion, deletion and site-directed mutagenesis. The recombinant proteins were purified from bacteria via DEAE A-50 and Sephacryl S-200 chromatography, and their survival promoting activities were determined using cultures of embryonic chick dorsal root ganglion (DRG) neurons. Insertion at position 23 with APGL, or at position 79 with PRGA, or substitution of 162L163Q for PIDG resulted in proteins with no neurotrophic activity. However, insertion at position 186 with PRGI did not alter human CNTF activity. Deletion of the carboxy-terminal amino acid 186-200 did not reduce the biological activity, but elimination of the amino acid 162-186 abolished the activity. The mutant substituting of 17 Cys for Ser was found to display a biological activity equivalent to that of the wild type. Our data provided experimental confirmation for the structural prediction of CNTF.

Amino Acid Sequence

Regulation of glibenclamide receptors in cultured chick cardiomyocytes.

We found that the expression of sulphonylurea receptors which control ATP-dependent potassium channels was up-regulated both by a receptor agonist and by potassium channel activators. Sulphonylurea receptors detected on cell membranes of cultured chick cardiomyocytes by [3H]glibenclamide were increased 2.8-fold following growth of the cells for 1-2 days in the presence of 30 nM glibenclamide. [3H]glibenclamide binding was also increased approximately 2-fold following growth of the cells in the presence of the ATP-dependent potassium channel activators, pinacidil, minoxidil and diazoxide, but was diminished to 75% of control following treatment with depolarizing concentrations of KCl.

Animals

Expression of leukemia inhibitory factor and interleukin-11 by human melanoma cell lines: LIF, IL-6, and IL-11 are not coregulated.

Dysregulation in cytokines has been associated with melanomas. For example, loss of growth inhibition in advanced melanomas has been associated with interleukin-6 (IL-6) expression. Because IL-6 belongs to the hematopoietic cytokine family, which includes leukemia inhibitory factor (LIF) and interleukin-11 (IL-11), we examined the possibility of coordinate expression of LIF, IL-6, and IL-11 in three human melanoma cell lines derived from primary lesions (early) and in four lines derived from metastatic tumors (advanced). All lines examined produced at least low levels of LIF and IL-11 mRNA as measured by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). By enzyme-linked immunosorbent assay (ELISA), two of three early and three of four advanced lines were found to secrete LIF protein. IL-11 was assayed using growth of the responsive B9/11 cell line, but only one of seven lines made a low but measurable amount of IL-11. Cytokine protein production was not strictly correlated with mRNA abundance, nor was it strongly correlated with tumor staging. Recombinant LIF and IL-11 protein had no effect on the proliferation of any of the seven lines, suggesting that they do not act as autocrine growth factors for these melanomas. Assay of IL-6, IL-11, and LIF protein in conditioned medium from early and advanced melanoma lines gave no evidence of coordinate expression of these cytokines. We conclude that LIF and IL-11 production by melanomas may have some paracrine or endocrine function in the course of melanoma progression.

Base Sequence

[Repair of scalp defects with skull exposure].

The methods for repairing scalp defects and skull exposure are introduced. From 1980 to 1991, 9 cases of scalp defects and skull exposure from injury or tumor extirpation were treated with free omentum transfer with split skin graft, free flap or axial flap transfer. The surgical procedure was described in detail. For the whole defect of scalp with skull exposure the authors consider the free anterior-lateral femoral flap more appropriate and suggest that the free omentum flap be avoided as far as possible.

Adolescent

[Diagnosis and treatment of primary ureteral carcinoma--a report of 12 cases].

Twelve cases with primary ureteral carcinoma are reported with a review of literatures. Urography was the main method of diagnosis, and retrograde uretero-pyelography was more helpful. Cystoscopy was mandatory before operation. CT was useful for diagnosis and staging. If needed, ureteroscopy could be used to increase diagnostic accuracy. The traditional total nephroureterectomy with a cuff excision of the urinary bladder was the first and basic choice of treatment. Regular cystoscopy, excretory urography and adjuvant intravesical therapy should be carried out after operation.

Aged

[Application of T-subgroup cells assay in the immunology of oral-maxillofacial cancer].

We report on T-subgroup cells in 70 cases of oral-maxillofacial cancer determined by T-monoclone antibody-OKT system. The results are as follows: CD3, CD4/CD8 value of cancer patients was decreased significantly, compared with that of health or oral benign tumor patients. CD8 was significantly increased (P < 0.001); however, CD8 was decreased and CD4/CD8 ratio was increased after operation. 20 cases underwent the dynamic testing, determined in 1 week, 1 and 3 month. The results are as follows: CD3, CD4, CD4/CD8 values were increased gradually and CD8 was decreased gradually in 17 cases of curative patients, but CD4, CD4/CD8, values were decreased and CD8 was increased significantly in 3 cases of metastased patients. Our conclusion is that determination of T-subgroup cells is of reference value for monitoring and treating cancer patients.

Adenocarcinoma