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Biomedical subjects

H Lee

Publications and source records attributed to H Lee.

At least 361 records · Page 20Linked to original sources

Cold-regulated gene expression and freezing tolerance in an Arabidopsis thaliana mutant.

Low temperature is an important environmental factor influencing plant growth and development. In this study, we report the characterization of a genetic locus, HOS2, which is defined by three Arabidopsis thaliana mutants. The hos2-1, hos2-2 and hos2-3 mutations result in enhanced expression of RD29A and other stress genes under low temperature treatment. Gene expression in response to osmotic stress or ABA is not affected in the hos2 mutants. Genetic analysis indicates that the hos2 mutations are recessive and in a nuclear gene. Compared with the wild-type plants, the hos2-1 mutant plants are less capable of developing freezing tolerance when treated with low non-freezing temperatures. However, the hos2-1 mutation does not impair the vernalization response. These results indicate that HOS2 is a negative regulator of low temperature signal transduction important for plant cold acclimation.

Acclimatization↗

HOS5-a negative regulator of osmotic stress-induced gene expression in Arabidopsis thaliana.

Osmotic stress activates the expression of many plant genes through ABA-dependent as well as ABA-independent signaling pathways. We report here the characterization of a novel mutant of Arabidopsis thaliana, hos5-1, which exhibits increased expression of the osmotic stress responsive RD29A gene. The expression of several other stress genes are also enhanced by the hos5-1 mutation. The enhanced expression is specific to ABA and osmotic stress because low temperature regulation of these genes is not altered in the mutant. Genetic analysis indicated that hos5-1 is a recessive mutation in a single nuclear gene on chromosome III. Double mutant analysis of hos5-1 and the ABA-deficient aba1-1 as well as the ABA-insensitive abi1-1 mutant indicated that the osmotic stress hypersensitivity of hos5-1 is not affected by ABA deficiency or insensitivity. Furthermore, combined treatments of hos5-1 with ABA and osmotic stress had an additive effect on RD29A-LUC expression. These results suggest that the osmotic stress hypersensitivity in hos5-1 may be ABA-independent. The germination of hos5-1 seeds was more resistant to ABA. However, the hos5-1 mutation did not influence stomatal control and only slightly affected the regulation of growth and proline accumulation by ABA. The hos5-1 mutation reveals a negative regulator of osmotic stress-responsive gene expression shared by ABA-dependent and ABA-independent osmotic stress signaling pathways.

Arabidopsis↗

Panax ginseng protects the testis against 2,3,7, 8-tetrachlorodibenzo-p-dioxin induced testicular damage in guinea pigs.

OBJECTIVES: To investigate histopathologically the beneficial effects of Panax ginseng extract on guinea pig testes damaged by 2,3, 7,8-tetrachlorodibenzo-p-dioxin (TCDD). MATERIALS AND METHODS: Ninety guinea pigs were divided into six equal groups. The normal controls (group 1) received vehicle and saline; group 2 received TCDD (1 microgram/kg) intraperitoneally; group 3 and 4 received 100 or 200 mg/kg per day of Panax ginseng water extract (PG-WE) intraperitoneally for 28 days from 1 week before TCDD injection; groups 5 and 6 received PG-WE for 14 days from 1 week after TCDD treatment. RESULTS: The gain in body weight was less in groups treated with TCDD than in controls. Moreover, the body weight of group 2 decreased from 14 days after TCDD exposure, while that of groups 3 and 4 increased; there was no decrease in body weight in groups 3-6. The decrease in testicular weight caused by TCDD was prevented by PG-WE. Light microscopy showed smaller tubules and late maturation arrest in group 2; electron microscopy showed a dissolution of the germinal epithelium, disrupted tight junctions between adjacent Sertoli cells, and altered germ cells at all developmental stages. The maturation arrest in germ cells caused by TCDD was ameliorated in groups 3-6. The testes almost completely recovered in groups 3 and 4 and there was some therapeutic effect of PG-WE in groups 5 and 6. CONCLUSIONS: These results confirm the protective and therapeutic effects of Panax ginseng on atrophy and testicular damage induced by TCDD, providing evidence that ginseng might be a useful agent in preventing and treating testicular damage induced by environmental pollutants.

Animals↗

Sense of control and adjustment to breast cancer: the importance of balancing control coping styles.

The relationship of modes of control and desire for control to psychosocial adjustment in women with breast cancer was examined. Fifty-eight women with stage I or stage II breast cancer were surveyed shortly after their diagnosis and again 4 and 8 months later. The authors hypothesized that a control profile in which individuals use a positive yielding (i.e., accepting) mode of control in conjunction with an assertive mode results in better adjustment than relying exclusively or primarily on an assertive mode. Results lend preliminary support to this hypothesis. At 8-month follow-up, those women who had a high desire for control and were low in positive yielding control showed the poorest adjustment, whereas those high in desire and the positive yielding mode showed the best psychosocial adjustment. The findings suggest that balanced use of active and yielding control efforts may lead to optimal psychosocial adjustment and quality of life in the face of life-threatening illnesses.

Adaptation, Psychological↗

Stress and course of disease in multiple sclerosis.

In this prospective study, 96 healthy controls and 101 multiple sclerosis patients were followed up for as many as 6 years, and self-reported stressful events and health status were assessed. The authors evaluated (a) whether patients reported more stressful life events than healthy controls and (b) the bidirectional relationship between stress and functional deterioration among patients. Healthy controls reported more life events than patients, Odds ratio (OR) = 1.13, p < .0001; and this relationship was attributable to healthy controls' reporting more neutral/positive events than patients. A bidirectional relationship was confirmed between stress and illness: there was an increased risk of disease progression when rate of reported stressful events was higher, OR = 1.13, p < .0003, and an increased risk of reported stressful events when rate of disease progression was higher, OR = 2.13, p < .0001. There were no differences in reported stress by level of baseline disability. The authors concluded that multiple sclerosis patients demonstrate a vicious cycle between stress and disease progression.

Adaptation, Psychological↗

Potentiation of cholecystokinin and secretin-induced pancreatic exocrine secretion by endogenous insulin in humans.

To investigate the effects of endogenous insulin on pancreatic exocrine secretion in humans, we evaluated the pure pancreatic juice obtained by endoscopic cannulation of the main pancreatic duct in 21 healthy subjects (14 men and seven women). Samples of pancreatic juices were collected after intravenous injection of either glucose (50%, 40 ml), secretin (0.25 CU/kg), and cholecystokinin-8 (CCK) (40 ng/kg), or a combination of glucose, secretin, and CCK in six 5-min periods. The responses of plasma glucose, insulin, and C-peptide to intravenous administration of glucose were measured. After infusion of glucose, the plasma insulin and C-peptide levels were significantly increased and remained at high levels during 30-min experiments, intravenous administration of secretin and CCK resulted in significant increases of pancreatic secretion including volume, bicarbonate, and protein output. When glucose was simultaneously administered with secretin and CCK, pancreatic secretion was significantly increased, more than the effects achieved by the secretin and CCK. However, glucose alone did not increase basal pancreatic secretion. These observations suggest that endogenous insulin intensifies pancreatic secretion stimulated by secretin and CCK in humans.

Adult↗

ECG data compression using cut and align beats approach and 2-D transforms.

A new electrocardiogram (ECG) data compression method is presented which employs a two dimensional (2-D) transform. This 2-D transform method utilizes the fact that ECG signals generally show two types of redundancies--between adjacent heartbeats and between adjacent samples. A heartbeat data sequence is cut and beat-aligned to form a 2-D data array. Any 2-D compression method can then be applied. Transform coding using the 2-D discrete cosine transform (DCT) [2-D DCT] is employed here as an example. Using selections from the MIT-BIH arrhythmia and Medtronic databases, results are presented that illustrate substantial improvement in compression ratio over one-dimensional methods for comparable percent root-mean-square difference (PRD).

Algorithms↗

Specific detection of the gene for the extracellular neutral protease of Bacillus cereus by PCR and blot hybridization.

A pair of primers and a gene probe for the amplification and detection of the Bacillus cereus neutral protease gene (NPRC) were developed. Specificity for the npr genes of the B. cereus group members B. cereus, B. mycoides, and B. thuringiensis was shown. Restriction polymorphism patterns of the PCR products confirmed the presence of the NPRC gene in all three species.

Bacillus cereus↗

Mapping of the hepatitis B virus reverse transcriptase TP and RT domains by transcomplementation for nucleotide priming and by protein-protein interaction.

Hepadnavirus polymerases initiate reverse transcription in a protein-primed reaction. We previously described a complementation assay for analysis of the roles of the TP and RT domains of HBV reverse transcriptase (pol) in the priming reaction. Independently expressed TP and RT domains form a complex functional for in vitro priming reactions. To map the minimal functional TP and RT domains, we prepared baculoviruses expressing amino- and carboxyl-terminal deletions of both the TP and RT domains and analyzed the proteins for the ability to participate in transcomplementation for the priming reaction. The minimal TP domain spanned amino acids 20 to 175; however, very little activity was observed without a TP domain spanning amino acids 1 to 199. The minimal RT domain spanned amino acids 300 to 775; however, little activity was observed unless the carboxyl end of the RT domain extended to amino acid 800. Thus, most of the RNase H domain was required. In previous studies, we observed a TP inhibitory domain between amino acids 199 and 344. The current analysis narrowed this domain to residues 300 to 334, which is a portion of the minimal RT domain. In addition, the ability of TP and RT deletion mutants to form stable TP-RT complexes was examined in coimmunoprecipitation assays. The minimal TP and RT domains capable of protein-protein interaction were considerably smaller than the domains required for functional interaction in the transcomplementation assays, and unlike priming activity, TP-RT interaction did not require the epsilon RNA stem-loop. These studies help to further define the complex protein-protein interactions required in HBV genome replication.

Animals↗

Role of cellular tumor necrosis factor receptor-associated factors in NF-kappaB activation and lymphocyte transformation by herpesvirus Saimiri STP.

The STP oncoproteins of the herpesvirus saimiri (HVS) subgroup A strain 11 and subgroup C strain 488 are now found to be stably associated with tumor necrosis factor receptor-associated factor (TRAF) 1, 2, or 3. Mutational analyses identified residues of PXQXT/S in STP-A11 as critical for TRAF association. In addition, a somewhat divergent region of STP-C488 is critical for TRAF association. Mutational analysis also revealed that STP-C488 induced NF-kappaB activation that was correlated with its ability to associate with TRAFs. The HVS STP-C488 P10-->R mutant was deficient in human T-lymphocyte transformation to interleukin-2-independent growth but showed wild-type phenotype for marmoset T-lymphocyte transformation in vitro and in vivo. The STP-C488 P10-->R mutant was also defective in Rat-1 fibroblast transformation, and fibroblast cell transformation was blocked by a TRAF2 dominant-negative mutant. These data implicate TRAFs in STP-C488-mediated transformation of human lymphocytes and rodent fibroblasts. Other factors are implicated in immortalization of common marmoset T lymphocytes and may also be critical in the transformation of human lymphocytes and rodent fibroblasts.

Amino Acid Sequence↗

A precipitation reaction found in patients with hepatitis C as a marker for the purification of virus-like particles.

Even after the molecular cloning of the hepatitis C virus (HCV), an HCV-specific precipitation reaction has not yet been identified. We attempted to develop a precipitation system exclusively for anti-HCV-positive sera as a first step in finding an HCV-specific antigen and HCV-associated particles. In some patients being in a final stage of different liver diseases, we found sera (179/132,761; designated 'a-CK') which specifically precipitated with anti-HCV- and HCV-RNA-positive sera (designated 'CK'). When CK-positive sera were searched for in patients with various liver diseases using standard a-CK-positive plasma, CK was detected in 420 (57.9%) of 726 anti-HCV-positive sera and in none of the 1,630 anti-HCV-negative ones. The nature of CK and a-CK has not been fully clarified yet; CK demonstrated inter-betagamma mobility, whereas a-CK showed beta-globulin mobility; CK was not detected in cryoprecipitate, but HCV RNA was present in precipitates of CK-positive plasma incubated with one that was a-CK positive. Transmission electron microscopy revealed two size ranges of particles in the precipitate of CK- and a-CK-positive plasmas, 23-38 nm and 48-65 nm. We have found a novel precipitation system which is exclusive to anti-HCV-positive sera and which specifically precipitates an HCV-RNA-containing serum fraction and particles. This system can be useful for the purification and characterization of the circulating particles. Furthermore, it may be a new approach to the nature of HCV-RNA-carrying material.

Centrifugation↗

Differential changes of cell cycle regulators and activities in kidneys during pre- and postnatal development.

The present study was designed to determine the changes of the cyclin/CDK (cyclin dependent kinase)/CKI (CDK inhibitors) system in kidneys during pre- and postnatal development. All protein levels of cyclins (cyclins D1, D3, E, A, B) and protein levels and activities of CDKs (CDK4, CDK2, cdc2) were high in kidneys during the prenatal period and decreased differently during the postnatal period. As the phosphorylated active form of cyclin D1 decreased, the dephosphorylated inactive form of cyclin D1 increased during the early postnatal development. While CDK4 activities decreased markedly, the activities of CDK2 and cdc2 decreased gradually during the early postnatal period. While the p21(CIP1) protein was barely detectable during the prenatal period, but was not detectable during the postnatal period, the protein level of p27(KIP1) was detectable during pre- and postnatal periods. These results indicate that the cyclin/CDK/CKI system is actively involved in the nephrogenesis during the prenatal period and is closely associated with the withdrawal of the renal cell cycle during the postnatal period.

Age Factors↗

Quercetin inhibits benzo[a]pyrene-induced DNA adducts in human Hep G2 cells by altering cytochrome P-450 1A1 gene expression.

Quercetin is one of the most abundant of the naturally occurring flavonoids. It has been estimated that about 25-50 mg of quercetin are consumed from the daily diet. The chemopreventive effect of quercetin on dietary carcinogen has been intensely studied in animal models; however, knowledge regarding the molecular mechanism is still limited. In this study, the human hepatoma Hep G2 cell line was used to investigate how quercetin prevents benzo[a]pyrene (B[a]P)-induced DNA adducts. The Hep G2 cells were treated with 10 microM B[a]P for 18 hours in the presence or absence of quercetin. The DNA adduct levels, evaluated by 32P postlabeling, decreased in a dose-dependent manner after treatment with quercetin. Cytochrome P-450 1A1 (CYP1A1) and glutathione S-transferase involvement have been well demonstrated in the modulation of B[a]P-induced DNA damage. From the assays of both enzyme activities, quercetin inhibits CYP1A1-linked ethoxyresorufin O-dealkylase activity more effectively than glutathione S-transferase activity. To elucidate the molecular mechanisms, reverse transcriptase-polymerase chain reaction and Western blot were used to evaluate whether the decrease in CYP1A1 enzyme activity by quercetin is mediated because of alterations of CYP1A1 transcription or mRNA stability. The results indicated that quercetin significantly inhibits B[a]P-induced CYP1A1 mRNA and protein expression. From these findings, we conclude that quercetin suppresses B[a]P-induced DNA damage in human Hep G2 cells by altering CYP1A1 gene expression. Thus we suggest that dietary quercetin may have a long-term preventive effect on chemical carcinogenesis, especially in people who eat a diet rich in fruits and vegetables.

Anticarcinogenic Agents↗

A short term (accelerated release) approach to evaluate peptide release from PLGA depot-formulations.

An accelerated method to evaluate peptide release from poly(dl-lactide-co-glycolide) (PLGA) depot formulations in short time is described. Peptide-loaded microspheres were made from hydrophilic 50:50 PLGA by a dispersion-solvent extraction technique, and peptide release was studied at 37 degrees C and at higher temperatures in various media. For all accelerated conditions, release was faster at temperatures above the glass transition, Tg, of the host polymer. Complete release of peptide from 8600 MW PLGA was achieved in 35 hours at 50 degrees C in buffered and nonbuffered media containing 0.5% polyvinyl alcohol (PVA). Type of release media and concentration of PVA influenced the release profiles. A PVA concentration of 0.1 to 0.5% was found to prevent aggregation of microspheres at higher temperatures, with an increase in release at the higher PVA concentration. Peptide release was associated with a reduction of pH of the releasing media and increased mass loss. Complete peptide release at pH 4 from 8.6 kd and 28 kd PLGA at 50 and 60 degrees C occurred within 30-40 hours and correlated well with the real-time release at 37 degrees C and pH 7.0. At the higher molecular weight, a slightly longer accelerated release time and higher temperature were required to correlate with the real-time release. The data suggest that by optimization of release conditions such as temperature, surfactant concentration, buffer component, and pH, an accelerated study could be employed to evaluate depot formulations for a given polymer type.

Buffers↗

Inhibition of hypothalamic GnRH secretion in the ewe by antigonadotropic decapeptide during the estrous cycle and nonbreeding season.

Previous experiments from our laboratory and others have shown that the peptide antigonadotropic decapeptide (AGD) has marked inhibitory effects on luteinizing hormone (LH) secretion in rats and ewes. The first objective of this study was to determine whether AGD inhibits LH secretion by regulating hypothalamic release of gonadotropin hormone (GnRH). AGD (200 microg in 200 microL of 0.3% bovine serum albumin [BSA] saline) or vehicle was infused into the lateral ventricle of ovariectomized (OVX) ewes with hypophyseal-portal cannulae, and GnRH secretion was monitored. The frequency of GnRH and LH pulses in AGD-treated ewes was significantly decreased (p < 0.05) but did not change in the control ewes. The second objective of this investigation was to evaluate changes in hypothalamic sensitivity to AGD in the ewe during the estrous cycle and nonbreeding season. During the estrous cycle, the effects of AGD on LH secretion were assessed following ovariectomy, during the metestrous, diestrous, and proestrous phases of the estrous cycle. The response to AGD during the estrous cycle was compared to its effect during the anestrous season. LH, cortisol, and prolactin (PRL) concentrations were assayed in peripheral blood samples obtained at 10-min intervals over a 6-h period prior to injection of either vehicle (200 microL of 0.3% BSA in 0.9% saline) or AGD (200 microg in 200 microL of vehicle), and for an additional 10 h following treatment. LH pulse frequency decreased after treatment with AGD (p < 0.05) at all times in OVX and intact ewes compared to vehicle-treated controls. During the anestrous season, AGD treatment was more effective in inhibiting LH pulse frequency than during the breeding season (p < 0.05). Furthermore, there was a significant increase (p < 0.05) in mean cortisol concentrations after AGD infusion in all AGD-treated groups compared to controls independent of season or reproductive status. PRL concentrations were also increased (p < 0.05) following treatment with AGD. These results suggest that inhibition of pulsatile LH release induced by AGD is modulated by alterations in frequency of hypothalamic discharges of GnRH. Furthermore, changes in the inhibitory actions of AGD may contribute to the seasonal regulation of hypothalamic GnRH secretion in the ewe.

Anestrus↗

Radiofrequency ablation of a right atriofascicular Mahaim fiber and two contralateral left free-wall accessory pathways.

We report a rare combination of a right atriofascicular Mahaim fiber and two left-sided atrioventricular accessory pathways in a 57-year-old female presenting with an antidromic atrioventricular reciprocating tachycardia. Radiofrequency ablation was first targeted at the left lateral accessory pathway that served as the retrograde limb of the tachycardia. After elimination of the left lateral pathway, a bystander left posterolateral pathway was detected, and it too was successfully ablated. Although no tachycardia was reinducible, the Mahaim pathway was ablated because of its short effective refractory period. A discrete Mahaim potential recorded at the right atrial free-wall successfully guided the ablation.

Bundle of His↗

Morphological and biochemical analysis of anti-nuclear matrix protein antibodies in human sera.

Autoimmune sera have been used in the diagnosis of autoimmune diseases as well as the analysis of nuclear substructures. In an attempt to study the biological characteristics of the nuclear matrix, we screened human sera using immunofluorescent staining and immunoblot. We detected antibodies against nuclear matrix (NM), a remnant nonchromatin protein compartment after the treatment of detergent, salt and nuclease, in 212 out of 284 tested sera (74.6%) by immunoblot. Peptides with molecular weights of 70 kDa, 50 kDa and 25 kDa were detected in the order of frequency. Clinical informations of 198 out of 212 cases were available and went as follows: 38 cases were autoimmune diseases, such as systemic lupus erythematosus and rheumatoid arthritis; 132 non-autoimmune and non-neoplastic diseases; 16 neoplastic diseases and 12 cases unclassified. The immunofluorescent staining intensity by anti-nuclear matrix protein (NMP) antibodies decreased variably, but fibrillogranular, speckled and nucleolar immunolocalization patterns were retained after in situ fractionation. Ku70 and La protein were detected by anti-NMP antibodies. Immunolocalization by anti-NMP antibodies indicates that the NMPs constitute a variety of characteristic nuclear substructures and may serve as autoantigens in diverse human diseases. In addition, the presence of Ku70 and La protein as NMPs suggests that the NM can be functionally active in association with DNA or RNA.

Antigens, Nuclear↗