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Biomedical subjects

L C Wu

Publications and source records attributed to L C Wu.

At least 73 records · Page 4Linked to original sources

Transgenic tobacco plants expressing the geminivirus BL1 protein exhibit symptoms of viral disease.

Bipartite geminiviruses, such as squash leaf curl virus (SqLCV), encode two movement proteins (MPs), BR1 and BL1, that are essential for viral movement in and subsequent infection of the host plant. To elucidate the biochemical functions of these MPs and define their respective contributions to viral infection, we have generated transgenic Nicotiana benthamiana plants expressing SqLCV BR1 and BL1. Transgenic plants expressing BR1 or a truncated BL1 were phenotypically indistinguishable from wild-type N. benthamiana. In contrast, transgenic plants expressing full-length BL1, alone or in combination with BR1, were strikingly abnormal both in their growth properties and phenotypic appearance, with leaves that were mosaic and curled under, thus mimicking typical SqLCV disease symptoms in this host. BL1 was localized to the cell wall and plasma membrane fractions, whereas BR1 was predominantly in the microsomal membrane fraction. These findings demonstrate that expression of BL1 in transgenic plants is sufficient to produce viral disease symptoms, and they further suggest that BL1 and BR1 carry out distinct and independent functions in viral movement.

Base Sequence↗

Unrelatedness of groups of yeasts within the Candida haemulonii complex.

Isoenzyme and protein profiles of clinical isolates initially identified as Candida haemulonii demonstrated the presence of two distinct groups. DNA relatedness studies with representative cultures confirmed the presence of two species. Physiological features that can be used to separate two groups within C. haemulonii are reported.

Candida↗

Quantitative analysis of vascular prostaglandin H synthase mRNA levels by competitive polymerase chain reaction.

Prostaglandin H synthase (PGHS) occupies a central position in the biosynthesis of prostaglandins, thromboxane, and prostacyclin. Expression of this enzyme is enhanced by inflammatory cytokines and phorbol esters. Because of the low abundance of PGHS, PGHS mRNA levels in tissues and cells were difficult to measure accurately. In this study, we quantified vascular PGHS-1 mRNA levels by a competitive polymerase chain reaction assay. The mean basal PGHS-1 level in cultured human umbilical vein endothelial cells (HUVECs) was 24.3 +/- 10.6 amol/microgram RNA. Treatment of the HUVECs with recombinant human interleukin-1 beta at 37 degrees C for 4 hours increased the mRNA to 49.1 +/- 9.1 amol/microgram RNA, a 2.1-fold increase. Treatment of the cells with phorbol 12-myristate 13-acetate at 37 degrees C for 4 hours caused a 1.7-fold increase in the mRNA level. This procedure was utilized to measure PGHS-1 mRNA levels in rabbit abdominal aorta, inferior vena cava, liver, kidney, and lung. The aortic mRNA level (9.2 +/- 1.9 amol/microgram RNA) was about 5 times higher than the inferior vena caval level (2.4 +/- 1.3 amol/microgram RNA) but only slightly higher than the level of other organs. These results reveal a significant difference in PGHS-1 mRNA levels between arterial and venous tissues. The synthase mRNA levels in organs such as liver, kidneys, and lungs do not differ significantly. The PCR assay should be valuable for studying the role of PGHS expression in disease states.

Animals↗

[Staffing levels and patient needs in the intensive care unit].

The current study sought to utilize a patient classification system to investigate staffing and patient needs along with nursing care distribution in our intensive care units. The study employed a factor type analysis to design a patient need checklist (for our six ICUs) in order to determine staff load and nursing requirements. Snapshot observations were also taken to survey the distribution of nursing care time. The results of the two methods provide an estimate of current staffing needs, they also show that there is no significant difference between our surgical and internal medicine wards. On average, direct care accounts for 40.1% of the time schedule; indirect care, 37.3%; related care, 6.0%; and individual time, 16.6%. The average patient grade lies between I and III; and workload index, between 4.4 and 11.5. The average nursing time per patient per shift is 2.88 hours, and the average workload per person is estimated at 5.7-5.8 hours. By comparing the number of personnel currently employed and the estimated number needed, we discovered that two units are understaffed, and three are overstaffed. The understaffed units are all surgical units; the overstaffed ones, internal medicine units. To conclude the study, we examined the nature and complexity of nursing duties in the hopes of returning non-nursing responsibilities to the proper medical organizations. Our ultimate goal is to realize the full potential and improve the quality of our nursing personnel.

Female↗

Quantitative autoradiographic analysis of muscarinic cholinergic and GABAA (benzodiazepine) receptors in the forebrain of rats flown on the Soviet Biosatellite COSMOS 2044.

The quantitative autoradiographic analysis of muscarinic cholinergic and GABAA (benzodiazepine) receptors was performed on selected regions of the cerebral cortex and striatum of rats flown in the Soviet Biosatellite COSMOS 2044. An age- and strain-matched synchronous ground-based control group was employed for comparison. Muscarinic cholinergic receptor density was found to be significantly lower in the striatum of the flight animals as compared with that in the synchronous control group. No significant differences between flight and synchronous control groups were found in the other regions examined. GABAA (benzodiazepine) receptors showed no significant differences between the flight and control groups in any of the regions sampled. Although additional studies are needed to reach definitive conclusions, the decrease in muscarinic cholinergic receptors observed in the striatum suggests spaceflight-related alterations in motor activity.

Animals↗

Cellular expression of tropoelastin mRNA splice variants.

The primary transcript of tropoelastin is alternatively spliced into multiple mRNAs. The pattern and frequency of exon splicing is developmentally regulated, but the cellular profile of isoform expression within and among elastic tissues is not known. We used splice-variant specific antisense oligomeric deoxyribonucleotide probes in an in situ hybridization assay to assess the distribution of cells undergoing specific alternative splicing of tropoelastin pre-mRNA in developing bovine elastic tissues. Antisense oligomers were synthesized to exon sequences that are not alternatively spliced (exon 36) and to sequences that become abutted after high frequency (exon 33) and low frequency (exons 13 and 14) alternative splicing. The specificity of these probes for tropoelastin splice variants was verified by Southern hybridization to tropoelastin cDNAs with known exon deletions, and their specificity for tropoelastin mRNA was demonstrated by Northern hybridization. In situ hybridization with [35S]-labeled oligomers on sections of bovine lobar pulmonary artery and other elastic tissues showed that all elastogenic cells produce multiple forms of tropoelastin mRNA. These observations suggest that the production of tropoelastin isoforms is common to all cells within an elastin tissue and that this multiplicity may not be involved in regional differences in elastic tissue architecture.

Animals↗

Diagnosis of venous leakage by 133Xe corporeal clearance after intracavernous injection of prostaglandin E1 in poorly responding patients.

133Xe corporeal clearance after intracavernous (IC) prostaglandin E1 (PGE-1) injection was assessed in detecting venous leakage (VL) in a group of impotent patients with poor clinical response to PGE-1 injection. 133Xe corporeal washouts were done in the flaccid state and at full erection or at 20 min after the IC injection of 20 micrograms PGE-1 in cases where no full erection occurred in sequence on two separate days. In each case, data were acquired in frame mode after intracorporeal injection on one side of the midline just behind the glans of 0.1 ml, namely, 1-2 mCi (34-74 MBq) 133Xe in saline for 20 min. A time-activity curve was generated from the region of interest (ROI) at the site of injection and a computer routine was used to calculate clearance half-time (T1/2) in min and flow rate (Q) in ml per 100 g tissue per min. The data of 20 patients with equivocal response to IC PGE-1 were analysed. Of them, 14 had venous leakage and six had no vascular impairment (NVI). Venous leakage was proved by Doppler analysis, cavernosography after PGE-1, and/or selective arteriography. Twelve of 14 patients with VL had enhanced 133Xe corporeal clearance after PGE-1 with a significant decrease in T1/2 (mean +/- S.D.) from 115.9 +/- 196.0 to 13.3 +/- 13.4 (P less than 0.05) and increase in Q from 2.1 +/- 2.2 to 7.5 +/- 5.6 (P less than 0.01). In contrast, all six patients with NVI had decreased 133Xe corporeal clearance.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Sequence-specific interaction with the viral AL1 protein identifies a geminivirus DNA replication origin.

The bipartite geminiviruses such as tomato golden mosaic virus (TGMV) and squash leaf curl virus (SqLCV) have two single-stranded circular genomic DNAs, the A and B components, thought to be replicated from double-stranded circular DNA intermediates. Although it has been presumed that the origin sequences for viral replication are located in the highly conserved 200-nucleotide common region (CR) present in both genomic components and that the viral-encoded AL1 protein interacts with these sequences to effect replication, there has been no evidence that this is in fact so. We have investigated these questions, demonstrating selectivity and sequence specificity in this protein-DNA interaction. Simple component switching between the DNAs of TGMV and SqLCV and analysis of replication in leaf discs showed that whereas the A components of both TGMV and SqLCV promote their own replication and that of their cognate B component, neither replicates the noncognate B component. Furthermore, using an in vivo functional replication assay, we found that cloned viral CR sequences function as a replication origin and direct the replication of nonviral sequences in the presence of AL1, with both circular single-stranded and double-stranded DNA being synthesized. Finally, by the creation of chimeric viral CRs and specific subfragments of the viral CR, we demonstrated sequence-specific recognition of the replication origin by the AL1 protein, thereby localizing the origin to an approximately 90-nucleotide segment in the AL1 proximal side of the CR that includes the conserved geminiviral stem-loop structure and approximately 60 nucleotides of 5' upstream sequence. By deletional analysis, we further demonstrated that the conserved stem-loop structure is essential for replication. These studies identify the functional viral origin of replication within the CR, demonstrating that sequence-specific recognition of this origin by the AL1 protein is required for replication.

Base Sequence↗

Strain delineation and epidemiology of Candida (Clavispora) lusitaniae.

Electrophoretic karyotype (EK) patterns, determined by using contour-clamped homogeneous pulsed-field electrophoresis, and isoenzyme (IZ) profiles were evaluated as methods for strain delineation among 35 isolates of Candida lusitaniae recovered from 15 patients. All isolates were identified to the species level by using conventional morphologic and physiologic criteria, and the identification was confirmed by gas-liquid chromatography analysis of the cellular fatty acids. The isolates were then typed without knowledge of the patient source. The IZ profiles showed all isolates to be closely related. Fifteen EK patterns were found; each pattern was restricted to isolates recovered from a single patient. In contrast, on the basis of heterogeneity in phosphatases, beta-glucosidases, esterases, and catalases, 10 IZ profiles were found; 4 were shared by isolates recovered from more than one patient. Multiple isolates from six patients were analyzed, and for each patient, a single EK- and IZ-defined type was found. The types of isolates obtained from two patients, after the emergence of resistance to amphotericin B, remained the same as the types of isolates obtained earlier. The data suggest that a patient becomes colonized by a single strain of C. lusitaniae which may disseminate to multiple sites, that the colonizing strain can persist during the patient's hospitalization, and that it may develop resistance to amphotericin B. Both EK patterns and IZ profiles can be used to delineate strains of C. lusitaniae, but the EK pattern provides more discriminatory power.

Amphotericin B↗

Expression and characterization of the trans-activating protein Tax of human T-cell leukemia virus type I in Saccharomyces cerevisiae.

The trans-activator protein Tax of human T-cell leukemia virus type I (HTLV-I) stimulates transcription of the viral genome from the long terminal repeat. With a reporter HIS4TATA::lacZ fusion gene, the transcriptional activity of the Tax-responsive element in the long terminal repeat was tested in Saccharomyces cerevisiae. We found that fragments containing the 21-bp repeat of the HTLV-I enhancer stimulate synthesis of beta-galactosidase activity 15- to 20-fold. To test the ability of the Tax protein to trans activate the HTLV-I enhancer in yeast cells, the pX region of HTLV-I, encoding the Tax protein, was cloned under the control of the yeast GAL1 promoter. The expressed Tax protein is localized in the nucleus and associated with the yeast nuclear matrix fraction. In yeast cells that contained the integrated tax gene, two- to sixfold stimulation of expression from the HTLV-I enhancer was detected at the early stages of tax induction. This in vivo reconstitution system provides a new approach for examining the host factor(s), the signal transduction mechanism(s), and the role of nuclear architecture involved in Tax-mediated trans activation.

Blotting, Western↗

Isoenzyme changes in Candida albicans during domestication.

Isoenzyme analysis was performed on multiple strains of two commonly used reference cultures of Candida albicans (B311 and NCPF3153). Whereas strains originating from C. albicans B311 showed no variation in isoenzyme profiles, some strains derived from NCPF3153 were identical to B311 strains but others showed variation in their glucose-6-phosphate dehydrogenase isoenzymes. The results are compared with those from previous analyses with these strains and show that C. albicans can undergo genetic alterations during prolonged maintenance in laboratories.

Candida albicans↗

[Studies of the blood-group of chicken. X. Analysis of blood-group and plasma protein polymorphism in eleven Chinese native fowl breeds].

This paper reports the results of blood-group and plasma protein polymorphism of eleven Chinese native fowl breeds. 1. The distribution frequencies of the blood-group genes (3 loci 12 alleles) in eleven Chinese native fowl breeds were significantly different in A and C loci, but not significant in B locus. The coefficients of homozygosity of blood-group gene in all breeds were almost similar, except in Gushi and Chonren pitted chickens. The distribution of blood-group factors in all breeds was extensive, and this means that the selection potential in these local breeds was very large. 2. The gene frequencies of alkaline phosphatase (Akp and Akp-2) transferrin (Tf) in some breeds were more different, while that of esterase (Es-1) was less different. Among Chinese Japanese and American-European native breeds, the difference of gene frequencies of esterase was greater, while that of alkaline phosphatase and transferrin was smaller. It showed the identity or diversity in breed origin and evolution in these Chinese local breeds and Japanese or American-European native breeds. 3. The cluster analysis of the eleven Chinese native fowl breeds showed that these breeds could be divided into four groups: White ear-lobe- Shouguang- Luyuan chickens; Xiaoshan-Xianju- Pudong chickens; Langshan- Taihe Silky- Beijing Youkei chickens and Gushi-Chonren pitted chickens.

Alkaline Phosphatase↗

Radionuclide imaging clinical study. Corporeal 133Xe washout for detecting venous leakage in impotence.

A quantitative test, corporeal 133Xe washout, was developed to detect venogenic impotence. Data were acquired in frame mode after intracorporeal injection of 0.1 ml (1-2 mCi or 37-74 MBq) of 133Xe in saline on one side of the midline just behind the glans in the flaccid state for 20 min. A time-activity curve (TAC) was generated from the region of interest (ROI) at the site of injection and a computer routine was then used to calculate clearance half-time (T1/2) in minutes and flow rate (Q) in ml/100 g tissue/min from corporeal 133Xe clearance. The data from 12 controls and 21 patients with venous leakage were analysed. In patients with venogenic impotence, T1/2 (mean +/- S.D.) was greatly increased (24.4 +/- 10.7 min vs 10.7 +/- 4.6 min in controls, P less than 0.005) whereas Q markedly decreased (2.4 +/- 1.3 ml/100 g tissue/min vs 5.4 +/- 2.3 ml/100 g tissue/min in controls, P less than 0.005). The sensitivity and specificity for detecting venous leakage at the optimal threshold of T1/2 equal to 13 min and Q equal to 3.7 ml/100 g tissue/min were 95 and 75%, respectively. The corporeal 133Xe clearance half-time and flow rate are two simple and clinically useful indices for detecting venous leakage.

Adult↗

Transfer function analysis of the ventricular function: a new method for calculating right ventricular ejection fraction.

The relationship between the ventricular transfer function and ejection fraction has been investigated by the routine procedure of first-pass radionuclide angiocardiography. Ejection fraction has been shown to equal 1-e(-b), where b is the ratio of the R-R interval over the mean transit time difference between ventricular and atrial time-activity curves. To evaluate the effect of region of interest (ROI) on the right ventricular ejection fraction (RVEF), the results of the transfer function analysis (TFA) technique using precise ROI, TFA using rectangular ROI, and the routine method were compared. Regression analyses among RVEFs obtained from the above ROI methods yielded good correlations. Reliable RVEFs have been obtained even in the case of an improper bolus injection. Thus, the TFA technique is a new, simple, and reliable method for calculating RVEF without needing to outline the right ventricle precisely.

Heart↗

Xenon-133 hepatic retention ratio: a useful index for fatty liver quantification.

Xenon-133 hepatic retention ratio was developed for quantifying fatty liver. Data were acquired in frame mode in the hepatic region and both lung bases for 5 min after rebreathing 20 mCi of gaseous 133Xe and for another 5 min during washout. Static [99mTc]sulfur colloid liver imaging was performed with the patient in the identical position immediately after the ventilation study and data were stored for liver localization. A hepatic time-activity curve corrected for background activity was generated. The 133Xe retention ratio was derived by dividing the activity at 3.5 min after washout by the peak activity. The data of 16 controls and 20 patients with fatty liver were analyzed. The retention ratio (mean +/- s.d.) was greatly increased in patients with fatty infiltration (0.43 +/- 0.20 vs. 0.04 +/- 0.08 in controls, p less than 0.001). There was a strong positive correlation between the 133Xe retention ratios and percentage of fat on biopsy as assessed by the amount of the liver tissue occupied by fat globules on H & E stained sections. The 133Xe hepatic retention ratio is a simple, accurate and clinically useful index of detecting, quantifying and managing fatty infiltration of the liver.

Adult↗

Developmental regulation of tropoelastin isoforms.

The production of tropoelastin isoforms was examined by cell-free translation of nuchal ligament RNA from cows of various ages ranging from 110 days of gestation to adult. Tropoelastin polypeptides synthesized in vitro were immunoprecipitated with a specific monoclonal antibody and separated by high resolution sodium dodecyl sulfate-polyacrylamide gel electrophoresis. From these experiments three distinct tropoelastin isoform patterns were detected at different developmental periods. 1) The fetal period was characterized by three polypeptides of about 67, 65, and 63 kDa produced in a ratio of 1:3:2. 2) In neonatal tissue, the same molecular mass forms were detected; however, the relative isoform ratio changed to 2:3:1. 3) From adult ligament RNA, only the 67- and 65-kDa forms of tropoelastin were detected, and these were produced in equal amounts. The pattern of tropoelastin isoforms produced from RNA isolated from other fetal elastic tissues was the same as for fetal ligament. Additional studies demonstrated that the relative amount of functional tropoelastin mRNA correlated with steady state levels of mRNA consistent with transcriptional regulation of tropoelastin synthesis. Although the significance of developmental switching of individual tropoelastin isoforms is unknown, it is likely that this process is required for proper assembly and, thus, function of elastic tissue.

Aging↗

Terminal differentiation of nuchal ligament fibroblasts: characterization of synthetic properties and responsiveness to external stimuli.

The temporal expression of elastogenesis is unique among connective tissues in that elastin production occurs primarily during late fetal and early neonatal periods and is essentially fully repressed once fiber assembly is completed. To test whether elastin synthesis in adult nuchal ligament fibroblasts is permanently repressed or whether the cells retain the ability to reinitiate production upon proper stimulation, we examined in adult ligament cells various parameters known to be involved in the regulation of elastin production. Elastin synthetic capacity, as determined by the levels of steady-state tropoelastin mRNA, of adult tissue was significantly decreased relative to fetal tissue. Likewise, fibroblasts grown from explants of adult ligament had about a fourfold decrease in elastin production and elastin-specific mRNA levels. On the other hand, adult cells were similar to fetal ligament cells in that they were sensitive to glucocorticoid stimulation and demonstrated chemotactic responsiveness to elastin peptides. Since our previous studies have shown that the extracellular matrix (ECM) plays an important role in influencing elastin phenotypic expression, fetal and adult fibroblasts were grown on slices of nonviable adult ligament to test if repression of elastin production was directed by factors in ECM of adult tissues. No change in elastin synthesis was detected with either cell type grown on adult ligament, whereas both fetal and adult cells demonstrated increased elastin production in response to contact with fetal ligament. These results suggest that adult ligament ECM does not provide a metabolic signal to shut off the elastin gene and that adult cells remain responsive to external stimuli that may reinitiate high levels of elastin synthesis.

Animals↗