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Biomedical subjects

J Luo

Publications and source records attributed to J Luo.

At least 127 records · Page 7Linked to original sources

Aberrant SP-B mRNA in lung tissue of patients with congenital alveolar proteinosis (CAP).

Mutations in the surfactant protein (SP)-B gene are responsible for SP-B deficiency in congenital alveolar proteinosis (CAP) (Nogee et al. J Clin Invest 1994: 93: 1860-1883; Lin et al. Mol Genet Metab 1998: 64: 25-35; Klein et al. Pediatrics 1998: 132: 244-248; Ballard et al. Pediatrics 1995: 96: 1046-1052). The multigenerational consanguineous pedigree under study does not carry any of the known mutations, although this pedigree had 14 infant deaths following respiratory distress at birth. Immunostaining of the lungs from three such infants revealed decreased or absent SP-B. By sequencing of SP-B exons, exon-intron junctions, and the 5' and 3' flanking regions, nine polymorphisms were found in this pedigree, but none of them could explain the observed SP-B deficiency. Further analysis of SP-B mRNA by reverse transcription-polymerase chain reaction from paraffin-embedded lung tissue of CAP patients showed that SP-B mRNA is not intact. Although the sequence of mRNA from exon 1-exon 7 and from exon 8-exon 10 could be amplified, the region between exons 7 and 8 could not. From fluorescence in situ hybridization of the short arm of chromosome 2p, only 2 signals were identified, eliminating the possibility of translocation as the cause of the SP-B mRNA aberrance. Although the nature of the genetic basis of SP-B deficiency in this family is currently unknown, the existence of aberrant SP-B mRNA may, at least in part, be responsible for the SP-B deficiency in this pedigree.

Chromosomes, Human, Pair 2↗

Polymorphisms of human SP-A, SP-B, and SP-D genes: association of SP-B Thr131Ile with ARDS.

An allele association study of 19 polymorphisms in surfactant proteins SP-A1, SP-A2, SP-B, and SP-D genes in acute respiratory distress syndrome (ARDS) was carried out. Trend-test analysis revealed differences (p < 0.05) in the frequency of alleles for some of the microsatellite markers flanking SP-B, and for one polymorphism (C/T) at nucleotide 1580 [C/T (1580)], within codon 131 (Thr131Ile) of the SP-B gene. The latter determines the presence or absence of a potential N-linked glycosylation site. Multivariate analysis revealed significant differences only for the C/T (1580) polymorphism. When the ARDS population was divided into subgroups, idiopathic (i.e., pneumonia, etc.) or exogenic (i.e., trauma, etc.), significant differences were observed for the C/T (1580), for the idiopathic ARDS group, and the frequency of the C/C genotype was increased in this group. Based on the odds ratio, the C allele may be viewed as a susceptibility factor for ARDS. Although the expression of both C and T alleles occurs in heterozygous individuals, it is currently not known whether these alleles correspond to similar levels of SP-B protein. These data suggest that SP-B or a linked gene contributes to susceptibility to ARDS.

Alleles↗

The function of PML in p53-dependent apoptosis.

The PML gene of acute promyelocytic leukaemia (APL) encodes a growth- and tumour-suppresor protein that is essential for several apoptotic signals. The mechanisms by which PML exerts its pro-apoptotic function are still unknown. Here we show that PML acts as a transcriptional co-activator with p53. PML physically interacts with p53 both in vitro and in vivo and co-localizes with p53 in the PML nuclear body (PML-NB). The co-activatory role of PML depends on its ability to localize in the PML-NB. p53-dependent, DNA-damage-induced apoptosis, transcriptional activation by p53, the DNA-binding ability of p53, and the induction of p53 target genes such as Bax and p21 upon gamma-irradiation are all impaired in PML-/- primary cells. These results define a new PML-dependent, p53-regulatory pathway for apoptosis and shed new light on the function of PML in tumour suppression.

Animals↗

Inhibition of cell proliferation in HCC-9204 hepatoma cells by a c-myc specific ribozyme.

A ribozyme (RZ) gene targeting c-myc mRNA was synthesized and cloned. Cleavage reaction showed that cleavage of the RZ was efficient and specific. The RZ gene-containing retrovirus vector pDOR-RZ was transfected into HCC-9204 hepatoma cells, which constitutively express high levels of c-myc using Lipofectamine. Positively transfected cells were selected using G418. In situ hybridization showed that both pDOR-RZ and pDOR vectors had been integrated into the chromosome of HCC-9204 cells. Dot blot hybridization indicated that expression of the RZ was only evident in pDOR-RZ-transfected HCC-9204 cells. Avidin-biotin complex enzyme-linked immunosorbent assay showed that c-myc expression was down-regulated. Chromatin aggregation into compact masses, cytoplasmic vacuole degeneration, and blurring of cytoplasm structure were observed by transmission electron microscopy in HCC-9204-RZ cells. These results suggest that the use of a c-myc mRNA cleaving enzyme could be most effective in tumor cells that are highly proliferative and constitutively express high levels of c-myc.

Carcinoma, Hepatocellular↗

Antihyperglycemic acetylenic glucosides from Bidens pilosa.

In vivo bioassay-guided fractionation of the aqueous alcohol extract of the aerial parts of Bidens pilosa Sch. Bip. var. radiata (Asteraceae) using C57 BL/Ks-db/db mice as a model for type 2 diabetes, yielded two known polyacetylenic glucosides, identified as 2-beta-D-glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7,9,11-+ ++triyne (1) and 3-beta-D-glucopyranosyloxy-1-hydroxy-6(E)-tetradecene-8,10,1 2-triyne (2). A 3:2 mixture of compounds 1 and 2 effected a significant drop in blood glucose.

Acetylene↗

Central skull base invasion of maxillofacial tumors: computed tomography appearance.

OBJECTIVE: The aim of this study was to demonstrate computed tomography (CT) features of maxillofacial tumors invading the central skull base and to offer information in aiding diagnosis. METHODS: Fifty-eight maxillofacial tumors with central skull base invasion shown on axial and coronal CT images were divided into benign (13 cases) and malignant (45 cases) groups, based on their pathologic outcomes as proven by biopsy and surgery. RESULTS: Four manifestations of the skull base abnormality were observed on CT images: (1) resorption of central skull base (53 cases, 11 benign and 42 malignant tumors); (2) enlargement of the foramen and canal in the central skull base (20 cases, 4 benign and 16 malignant tumors); (3) thinning of the central skull base (11 cases, 8 benign and 3 malignant tumors); and (4) displacement of the central skull base (5 cases, all benign tumors). The following structures of the central skull base were involved: the roof of the pterygoid process of the sphenoid bone (49 cases), the sphenoid greater wing (37 cases), the sphenoid body and sinus (33 cases), the petrous apex (12 cases), the clivus (4 cases), and the articular surface of the temporal squama (3 cases). Of 58 patients, 30 (7 benign and 23 malignant) had tumors with central skull base erosion that infiltrated into the cranial cavity. CONCLUSION: Benign maxillofacial tumors with center skull base erosion tended to lead to the displacement and thinning of the base of the middle skull fossa in contrast with tumors with malignant maxillofacial tumors. It is believed that these CT manifestations might be valuable in making a differential diagnosis.

Adolescent↗

Structural properties of hybrid triplex of polycation deoxyribonucleic S-methylthiourea (DNmt) strands with a complementary DNA strand, probed by nanosecond molecular dynamics.

The replacement of phosphodiester linkages of the polyanion DNA with S-methylthiourea linkers provides the polycation deoxyribonucleic S-methylthiourea (DNmt). Molecular dynamics studies to 1,220 ps of the hybrid triplex formed from octameric DNmt strands d(Tmt)8 with a complementary DNA oligomer strand d(Ap)8 have been carried out with explicit water solvent and Na+Cl- counterions under periodic boundary conditions using the CHARMM force field and the Ewald summation method. The Watson-Crick and Hoogsteen hydrogen-bonding patterns of the A/T tracts remained intact without any structural restraints for triplex structures throughout the simulation. The duplex portion of the triplex structure equilibrated at a B-DNA conformation in terms of the helical rise and other helical parameters. The dynamic structures of the DNmt x DNA x DNmt triplex were determined by examining histograms from the last 800 ps of the dynamics run. These included the hydrogen-bonding pattern (sequence recognition), three-centered bifurcating occurrences, minor groove width variations, and bending of tracts for the hybrid triplex structures. Together with the Watson-Crick hydrogen-bondings, the strong Hoogsteen hydrogen-bondings, the partially maintained three-centered bifurcatings in the Watson-Crick pair, and the medium-strength three-centered bifurcatings in the Hoogsteen pair suggest that the hybrid triplex is energetically favorable as compared to a duplex with similar base stacking, van der Waals interactions, and helical parameters. This is in agreement with our previously reported thermodynamic study, in which only triplex structures were observed in solution. The bending angle measured between the local axis vectors of the first and last helical axis segments is about 20 degrees for the Watson-Crick portion of the averaged structure. Propeller twist (associated with three-centered hydrogen-bonding) up to -30 degrees, native to DNA AT base pairing, was also observed for the triplex structure. The sugar pseudorotation phase angles and the ring rotation angles for the DNA strand are within the C3'-endo domain and C2'-endo domain for the DNmt strand. Water spines are observed in both minor and major grooves throughout the dynamics run. The molecular dynamics simulations of the structural properties of DNmt x DNA x DNmt hybrid triplex is compared to the DNG x DNA x DNG hybrid triplex (In DNG the -O-(PO2-)-O- linkers in DNA is replaced by -NH-C(=N+H2)-NH-).

Base Pairing↗

Association of the antidiabetic effects of bromocriptine with a shift in the daily rhythm of monoamine metabolism within the suprachiasmatic nuclei of the Syrian hamster.

Bromocriptine, a dopamine D2 agonist, inhibits seasonal fattening and improves seasonal insulin resistance in Syrian hamsters. Alterations in daily rhythms of neuroendocrine activities are involved in the regulation of seasonal metabolic changes. Changes in circadian neuroendocrine activities that regulate metabolism are believed to be modulated by central circadian oscillators within the hypothalamic suprachiasmatic nuclei (SCN) of seasonal animals. We examined the association of metabolic responses to bromocriptine with its effects on the daily rhythms of metabolic hormones and daily monoamine profiles within the SCN, a primary circadian pacemaker known to regulate metabolism, in Syrian hamsters. Obese glucose-intolerant male Syrian hamsters (body weight [BW] 185 +/- 10 g) held on 14h daily photoperiods were treated at light onset with bromocriptine (800 microg/animal/day, ip) or vehicle for 2 weeks. Animals were then subjected to a glucose tolerance test (GTT) (3 g/kg BW, ip). Different subsets of animals (n = 6) from each treatment group were sacrificed at 0h/24h, 5h, 10h, 15h, or 20h after light onset for analyses of SCN monoamines, plasma insulin, prolactin, cortisol, thyroxin (T4), triiodothyronine (T3), glucose, and free fatty acids (FFAs). Compared with control values, bromocriptine treatment significantly reduced weight gain (14.9 vs. -2.9 g, p < .01) and the areas under the GTT glucose and insulin curves by 29% and 48%, respectively (p < .05). Basal plasma insulin concentration was markedly reduced throughout the day in bromocriptine-treated animals without influencing plasma glucose levels. Bromocriptine reduced the daily peak in FFA by 26% during the late light span (p < .05). Bromocriptine significantly shifted the daily plasma cortisol peak from the early dark to the light period of the day, reduced the plasma prolactin (mean 1.8 vs. 39.4 ng/dL) and T4 throughout the day (mean 1.6 vs. 3.8 microg/dL), and selectively reduced T3 during the dark period of the day (p < .01). Concurrently, bromocriptine treatment significantly reduced SCN dopamine turnover during the light period and shifted daily peaks of SCN serotonin and 5-hydroxy-indoleacetic acid (5-HIAA) content by 12h from the light to the dark period of the day (p < .05). This was confirmed by a further in vivo microdialysis study in which bromocriptine increased SCN extracellular 5-HIAA of glucose-intolerant hamsters during the dark phase (47% increase, p < .05) toward levels observed in normal glucose-tolerant hamsters. Thus, bromocriptine-induced resetting of daily patterns of SCN neurotransmitter metabolism is associated with the effects of bromocriptine on attenuation of the obese insulin-resistant and glucose-intolerant condition. A large body of corroborating evidence suggests that such bromocriptine-induced changes in SCN monoamine metabolism may be functional in its effects on metabolism.

Animals↗

Chronic consumption of fresh but not heated yogurt improves breath-hydrogen status and short-chain fatty acid profiles: a controlled study in healthy men with or without lactose maldigestion.

BACKGROUND: Ingestion of fermented dairy products induces changes in the equilibrium and metabolism of the intestinal microflora and may thus have beneficial effects on the host. OBJECTIVE: We compared the effects of chronic consumption of yogurt with (fresh) or without (heated) live bacterial cultures (Lactobacillus bulgaricus and Streptococcus thermophilus) on plasma glucose, insulin, triacylglycerols, cholesterol, fatty acids, and short-chain fatty acids. DESIGN: Two groups of 12 healthy men with or without lactose malabsorption were selected with use of a breath-hydrogen test after a 30-g lactose load. Subjects were randomly assigned in a crossover design to 500 g/d of either fresh or heated yogurt for 2 periods of 15 d each, separated by a 15-d washout interval. RESULTS: Chronic consumption of fresh or heated yogurt had no detrimental effects on plasma glucose, insulin, or fatty acid areas under the curve in response to acute ingestion of 500 g yogurt in healthy men with or without lactose malabsorption. There were also no detectable changes in fasting plasma glucose, insulin, fatty acid, triacylglycerol, or cholesterol concentrations. In contrast, plasma butyrate was higher (P: < 0.03) and plasma propionate tended to be higher (P: = 0.059) in subjects without lactose malabsorption after fresh yogurt consumption than after heated yogurt consumption. There were no significant changes in plasma acetate. In subjects with lactose malabsorption, 15 d of fresh yogurt consumption also increased propionate production compared with values at baseline (P: < 0.04). In the same group, the production of breath hydrogen was lower after fresh yogurt consumption than after heated yogurt consumption (P: < 0.01). CONCLUSIONS: In men with lactose malabsorption, chronic consumption of yogurt containing live bacterial cultures ameliorated the malabsorption, as evidenced by lower breath-hydrogen excretion, but increased propionate concentrations. In subjects without lactose malabsorption, such yogurt tended to increase propionate and increased butyrate.

Adult↗

Chronic consumption of short-chain fructooligosaccharides does not affect basal hepatic glucose production or insulin resistance in type 2 diabetics.

Short-chain fructooligosaccharides (FOS) are prebiotics, which escape digestion in the small intestine and are fermented by the colonic microflora into short-chain fatty acids. Recently, we found that the daily consumption of 20 g FOS decreased basal hepatic glucose production in healthy subjects without any effect on insulin-stimulated glucose metabolism. In this study, we evaluated the effects of the chronic ingestion of FOS on plasma lipid and glucose concentrations, hepatic glucose production and insulin resistance in type 2 diabetics. Type 2 diabetic volunteers (n = 10; 6 men, 4 women) received either 20 g/d FOS or sucrose for 4 wk in a double-blind crossover design. FOS did not modify fasting plasma glucose and insulin concentrations or basal hepatic glucose production. The plasma glucose response to a fixed exogenous insulin bolus did not differ at the end of the two periods. Erythrocyte insulin binding also did not differ. Serum triacylglycerol, total and HDL cholesterol, free fatty acid, apolipoproteins A1 and B and lipoprotein (a) concentrations were not modified by the chronic ingestion of FOS. We conclude that 4 wk of 20 g/d of FOS had no effect on glucose and lipid metabolism in type 2 diabetics.

Blood Glucose↗

Purification, characterization, and molecular cloning of the gene of a seed-specific antimicrobial protein from pokeweed.

A small cysteine-rich protein with antimicrobial activity was isolated from pokeweed (Phytolacca americana) seeds and purified to homogeneity. The protein inhibits the growth of several filamentous fungi and gram-positive bacteria. The protein was highly basic, with a pI higher than 10. The entire amino acid sequence of the protein was determined to be homologous to antimicrobial protein (AMP) from Mirabilis jalapa. The cDNA encoding the P. americana AMP (Pa-AMP-1) and chromosomal DNA containing the gene were cloned and sequenced. The deduced amino acid sequence shows the presence of a signal peptide at the amino terminus, suggesting that the protein is synthesized as a preprotein and secreted outside the cells. The chromosomal gene shows the presence of an intron located within the region encoding the signal peptide. Southern hybridization showed that there was small gene family encoding Pa-AMP. Immunoblotting showed that Pa-AMP-1 was only present in seeds, and was absent in roots, leaves, and stems. The Pa-AMP-1 protein was secreted into the environment of the seeds during germination, and may create an inhibitory zone against soil-borne microorganisms. The disulfide bridges of Pa-AMP-1 were identified. The three-dimensional modeling of Pa-AMP-1 indicates that the protein has a small cystine-knot folding, a positive patch, and a hydrophobic patch.

Amino Acid Sequence↗

catena-Poly

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Journal Article↗

Role of protein phosphatases in the activation of CFTR (ABCC7) by genistein and bromotetramisole.

Genistein and bromotetramisole (Br-t) strongly activate cystic fibrosis transmembrane conductance regulator (CFTR; ABCC7) chloride channels on Chinese hamster ovary cells and human airway epithelial cells. We have examined the possible role of phosphatases in stimulation by these drugs using patch-clamp and biochemical methods. Genistein inhibited the spontaneous rundown of channel activity that occurs after membrane patches are excised from cAMP-stimulated cells but had no effect on purified protein phosphatase type 1 (PP1), PP2A, PP2B, PP2C, or endogenous phosphatases when assayed as [(32)P]PO(4) release from prelabeled casein, recombinant GST-R domain fusion protein, or immunoprecipitated full-length CFTR. Br-t also slowed rundown of CFTR channels, but, in marked contrast to genistein, it did inhibit all four protein phosphatases tested. Half-maximal inhibition of PP2A and PP2C was observed with 0.5 and 1.5 mM Br-t, respectively. Protein phosphatases were also sensitive to (+)-p-Br-t, a stereoisomer of Br-t that does not inhibit alkaline phosphatases. Br-t appeared to act exclusively through phosphatases since it did not affect CFTR channels in patches that had low apparent endogenous phosphatase activity (i.e., those lacking spontaneous rundown). We conclude that genistein and Br-t act through different mechanisms. Genistein stimulates CFTR without inhibiting phosphatases, whereas Br-t acts by inhibiting a membrane-associated protein phosphatase (probably PP2C) that presumably allows basal phosphorylation to accumulate.

Animals↗

Effects of mimosine on fiber shedding, follicle activity, and fiber regrowth in Spanish goats.

Ten 2-yr-old Spanish wethers (58.2 +/- 7.21 kg BW) were used to determine effects of 2-d intravenous infusion of mimosine (beginning on January 8) on fiber shedding, follicle activity, and fiber regrowth. Primary and secondary follicle activity on d 0 were 43 +/- 6.2% and 96 +/- 1.7%, respectively. Five wethers were infused with mimosine at 120 mg/(kg BW x d) and the other five received saline. At 7 to 10 d after the start of infusion, all five goats infused with mimosine exhibited shedding, whereas shedding by controls was not observed. Cashmere fiber shedding score (5-point scale: 1 = no shedding, 5 = excessive shedding) on d 4 was greater for mimosine goats than for controls (1.2 vs 2.0; P < .001), and shedding score for wethers receiving mimosine was greater (P < .05) on d 12, 16, and 20 than on d 0 and 4 (4.1 to 4.6 vs 1.4 and 2.0). Guard hair shedding score for goats receiving mimosine was greatest (P < .01) among the days after infusion for d 12 and greater (P < .01) on d 16 than on d 0 and 4. Nonetheless, cashmere fiber yield from combed fleece of mimosine goats (average of 73%) was much greater than for a clipping of the uncombed side (average of 28%) when the cashmere fiber shedding score exceeded 4.0. Secondary follicle activity on d 12 was lower (P < .01) for mimosine than for control wethers (6.8 vs 67.7%), and secondary follicle activity for mimosine-infused goats on d 12 was lower (P < .01) than on d 0 (98.9%), 4 (98.3%), and 20 (99.5%). Mimosine infusion resulted in no detectable fiber regrowth in wk 4 to 7 after the start of infusion, but regrowth rate in the following two 4-wk periods was similar for mimosine and control wethers. In conclusion, 2-d intravenous infusion of mimosine at 120 mg/(kg BW x d) in the winter induced cashmere shedding but had less effect on guard hairs, suggesting future potential use of chemicals such as mimosine to remove cashmere fiber.

Animals↗

[Study on biliary motility in cirrhotic patients with portal hypertension].

OBJECTIVE: To study biliary motility in cirrhotic patients with portal hypertension and analyze the relationship between these biliary motility and the degree of liver function. METHODS: Biliary motility function was detected in 40 patients with portal hypertension and 30 normal controls by using quantitative hepatobiliary scintigraphy. RESULTS: The rate of dysfunction of gallbladder in cirrhotic patients was 52.5%, and it was significantly higher than that in controls (chi(2)=11.92, P<0.01). Compared with the normal controls, the bile's transit in biliary duct of cirrhotic patients was slower (t=11.09, P<0.01), the time of gallbladder visualization became longer (t=5.65, P<0.01), and the gallbladder emptying time was markedly prolonged (t=2.10, P<0.05). A correlation was also seen between these biliary motility and the degree of liver function. CONCLUSION: Cirrhotic patients with portal hypertension often have abnormal motility of gallbladder and biliary duct.

Adult↗

Detection of mosaic chromosome 21 aneuploidy in vivo with CB-FISH method.

OBJECTIVE: To explore the mechanism by which the ratio of mosaic diploid cells in vivo increased in trisomy 21 cases. METHODS: Fluorescence in situ hybridization (FISH) with pericentric DNA probes specific to human chromosome 21 to the binucleated cells cytokinesis- blocked by cytochalasin-B(CB), namely, CB-FISH method. RESULTS: The mean frequency of diploid cells (1.69%+/-1.07%) was significantly higher than that of tetrasomy 21 cells(0.042%+/-0.064%)(P=0.000) in 12 trisomy 21 cases aged 0-9 years (mean 4 years) in vivo; whereas the incidence of spontaneous monosomy 21 cells (0. 265%+/-0.13%) was close to the frequency of trisomy 21 cells (0. 225%+/-0.184%) in 20 control individuals aged 0-10 years (mean 5 years). The frequency of cells undergone chromosome 21 nondisjunction was 0.822%+/-0.554% in cultured trisomy 21 cells, which was significantly higher than that in the cultured diploid cells (0.369%+/-0.25%). However, the rates of cells with chromosome 21 loss in the two cultured cell lines (0.01%+/-0.03% vs 0.043%+/-0.049%) were not significantly different. In both cultured cell lines, chromosome 21 nondisjunction occurred more frequently than loss. CONCLUSION: The increased frequency of diploidies in trisomy 21 cases may result from the accumulation of diploid cells originating from malsegregation of chromosome 21 in trisomic cells rather than the increase of chromosome 21 loss.

Aneuploidy↗

Endothelial cell migration onto metal stent surfaces under static and flow conditions.

Restenosis associated with intimal hyperplasia and thrombosis at sites of balloon angioplasty or stent placement remains an important clinical problem. It is likely that loss or damage to the arterial endothelium associated with these interventional procedures as well as the rate of its restoration plays a critical role in the extent of restenosis. Migration of arterial endothelial cells from adjacent intact endothelium is the predominant source of cells involved in re-endothelialization of the injured site. In this paper, we review the influence of hemodynamics on endothelial cell migration, both in vivo and in vitro. In addition, we present recent in vitro studies demonstrating the importance of the nature of metal substrates in modulating endothelial cell migration rate. Finally, we review the cellular and molecular mechanisms likely involved in governing endothelial cell migration, and relate them to a possible scenario of endothelial response to injury at sites of arterial intervention. Understanding the important factors regulating endothelial migration may provide insights that will ultimately lead to methods to accelerate endothelial healing and reduce the occurrence of arterial restenosis.

Angioplasty, Balloon, Coronary↗