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Biomedical subjects

H Chen

Publications and source records attributed to H Chen.

At least 235 records · Page 13Linked to original sources
PubMed2001

PARM: a practical utility for drug design.

To accommodate situations in which the 3D structure of the target receptor is not available, we have developed the Pseudo Atomic Receptor Model (PARM) software package. In this article we describe PARM and illustrate its use with three examples: elemenes (potential anticancer drugs), angiotensin converting enzyme inhibitors, and human HIV-1 inhibitors TTD (1,1,3-trioxo-2H, 4H-thieno[3,4-e][1,2,4] thiadiazine derivatives). The results show that PARM can build models with favorable cross-validation statistics (Rcv2 values 0.7-0.9) and give helpful SAR insight. PARM has certain advantages: (a) it can be used for many systems, regardless of whether the 3D structure of the receptor is known; (b) PARM models were demonstrated to be highly statistically reliable; and (c) PARM analyses are robust and reproducible.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed2001

Tailoring enzymes that modify nonribosomal peptides during and after chain elongation on NRPS assembly lines.

Nonribosomal peptide synthetases are large enzyme complexes that synthesize a variety of peptide natural products through a thiotemplated mechanism. Assembly of the peptides proceeds through amino acid loading, amide-bond formation and chain translocation, and finally thioester lysis to release the product. The final products are often heavily modified, however, through methylation, epimerization, hydroxylation, heterocyclization, oxidative cross-linking and attachment of sugars. These activities are the province of specialized enzymes (either embedded in the multidomain nonribosomal peptide synthetase structure or standalone).

Bacteria↗
PubMed2001

Sperm incorporation in Xenopus laevis: characterisation of morphological events and the role of microfilaments.

Scanning and transmission electron microscopy were used to determine the morphological changes in the egg plasma membrane associated with sperm binding, fusion and incorporation in Xenopus laevis. Sperm incorporation in Xenopus is rapid, occurring within 3-5 min following addition of sperm. Images have been obtained of both early sperm-egg interactions and fertilisation bodies. Additionally, two drugs that specifically alter F-actin dynamics, latrunculin and jasplakinolide, were used to determine whether sperm incorporation is a microfilament-dependent process. Jasplakinolide did not prevent sperm incorporation, cortical granule exocytosis or cortical contraction, suggesting these events can occur without depolymerisation of existing, stabilised filaments. Latrunculin A, which competes with thymosin beta4 in ooplasm for binding actin monomer, did not inhibit cortical granule exocytosis, but blocked cortical contraction in 100% of eggs at a concentration of 5 microM. Although a single penetrating sperm was found on an egg pretreated in latrunculin, fertilisation bodies were never observed. At < 5 microM latrunculin, many eggs did undergo cortical contraction with some exhibiting severe distortions of the plasma membrane and abnormal accumulations of pigment granules. Preincubation of eggs in jasplakinolide before latrunculin mitigated both these effects to some degree. However, eggs incubated in latrunculin either prior to or after insemination never progressed through first cleavage.

Actin Cytoskeleton↗
PubMed2001

Regulation of the PH-domain-containing tyrosine kinase Etk by focal adhesion kinase through the FERM domain.

Etk/BMX, a member of the Btk family of tyrosine kinases, is highly expressed in cells with great migratory potential, including endothelial cells and metastatic carcinoma cell lines. Here, we present evidence that Etk is involved in integrin signalling and promotes cell migration. The activation of Etk by extracellular matrix proteins is regulated by FAK through an interaction between the PH domain of Etk and the FERM domain of FAK. The lack of Etk activation by extracellular matrix in FAK-null cells could be restored by co-transfection with wild-type FAK. Disrupting the interaction between Etk and FAK diminished the cell migration promoted by either kinase. Furthermore, inhibiting Etk expression in metastatic carcinoma cell lines with an antisense oligonucleotide blocks integrin-mediated migration of these cells. Taken together, our data indicate the essential role of the interaction of the PH domain of Etk and the FERM domain of FAK in integrin signalling.

Animals↗
PubMed2001

NEDD4L on human chromosome 18q21 has multiple forms of transcripts and is a homologue of the mouse Nedd4-2 gene.

The validation of full-length cDNA represents a crucial step in gene identification and subsequent functional analysis. In searching for candidate genes for bipolar disorder on chromosome 18q21, a novel gene homologous to NEDD4 (Neural precursor cells expressed developmentally down-regulated) was identified using exon trapping and cDNA cloning. This novel gene is termed NEDD4L (Human Gene Nomenclature Committee symbol). Typical NEDD4 orthologues that contain a C2 (Ca(2+)/lipid-binding) and a HECT (Homologous to the E6-AP Carboxyl Terminus) ubiquitin-protein ligase domain, and multiple WW domains have been shown to regulate the epithelial sodium channel (ENaC). In mice, Nedd4 has two distinct isoforms termed Nedd4-1 that belongs to the typical NEDD4 class, and Nedd4-2 that is homologous to Nedd4-1 but lacks the C2 domain. NEDD4L contains the WW and HECT domains seen in the NEDD4 gene family, but lacks the C2 domain in the N-terminus. BLAST database search showed that the deduced polypeptide of NEDD4L has 97 and 62% sequence identity to mouse Nedd4-2 and human NEDD4, respectively. Multiple forms of transcripts of NEDD4L have been isolated, which differ in transcription start and termination sites together with the presence or absence of an alternative spliced exon. Northern blot analysis showed a 3.4 kb mRNA species was specifically expressed in heart and skeletal muscle, while a 3.2 kb band and/or an additional 3.6 kb band is seen in other tissues tested. Striking homology of NEDD4L to mouse Nedd4-2 suggests it is the human homologue of mouse Nedd4-2. Its position in a region of linkage for autosomal dominant orthostatic hypotensive disorder and its potential role in regulating ENaC make NEDD4L a candidate gene for this disorder.

Alternative Splicing↗
PubMed2001

Protection against ischemia/reperfusion injury and myocardial dysfunction by antisense-oligodeoxynucleotide directed at angiotensin-converting enzyme mRNA.

Angiotensin-converting enzyme (ACE) activity in the myocardium and angiotensin-II (Ang-II) levels in plasma increase after myocardial ischemia, which lead to exacerbation of myocardial injury and cardiac dysfunction. We examined the protective role of novel antisense-oligodeoxynucleotide (AS-ODN) directed at ACE mRNA in myocardial ischemic injury. Sprague-Dawley rats were treated with ACE-AS-ODN (200 microg per rat, n = 8, i.v.) or inverted-ODN (IN-ODN, 200 microg per rat, n = 8, i.v.), given with 600 microg per rat of liposome DOTAP/DOPE. Hearts from AS-ODN- or IN-ODN-treated rats were excised, perfused in vitro, and subjected to 25 min of global ischemia followed by 30 min of reperfusion. Parallel groups of rats were given ACE inhibitor captopril (5 mg/kg, n = 8) or saline (n = 8) before excising the hearts. Ischemia/reperfusion resulted in myocardial dysfunction (increase in coronary perfusion pressure and LV end-diastolic pressure and a decrease in developed LV pressure) in the saline-treated rats. Myocardial dysfunction was associated with evidence of lipid peroxidation and enzyme leakage (MDA and LDH levels in the myocardium) and up-regulation of ACE protein expression. Administration of AS-ODN or captopril, but not IN-ODN, reduced Ang-II levels in the plasma, decreased ischemia/reperfusion-mediated cardiac functional deterioration and lipid peroxidation, and preserved LDH in the myocardium (all P < 0.05 versus the saline group). AS-ODN and captopril had equipotent effects on cardiac dynamics. ACE protein expression (western blot) was decreased in the hearts of the AS-ODN-treated group, but not in IN-ODN-treated rat hearts. In contrast, ACE protein expression was significantly increased in captopril-treated rat hearts. These observations suggest that AS-ODN directed at ACE mRNA can ameliorate myocardial dysfunction and injury after ischemia/reperfusion, and its use is associated with decreased expression of ACE protein in the ischemic myocardium.

Angiotensin II↗
PubMed2001

P15INK4B gene methylation and expression in normal, myelodysplastic, and acute myelogenous leukemia cells and in the marrow cells of cured lymphoma patients.

P15INK4B methylation and expression was studied in bone marrow cells obtained from normal individuals, from patients who had been cured of lymphoma, and from patients with either MDS or AML. The level of p15 methylation was very low in normal BM cells and in CD34+ and CD34- subpopulations (0-6.5%; med, = 2.5%). P15INK4B transcripts were present in each of these cell populations. In contrast, methylation was the usual situation in MDS and AML marrows. The presence of methylation of the p15INK4B gene did not always indicate an absence of expression nor was expression always present if methylation was absent. P15INK4B methylation was studied in the marrows of nine patients (one studied twice) who had been cured of lymphoma and in whom hemopoiesis was believed to be normal. Increased methylaton was present in all 10 marrows. These data indicate that p15INK4B methylation is likely to be a very early event in the development of the secondary hematologic disorders.

Adult↗
PubMed2001

An acoustical study of the fricative /s/ in the speech of individuals with dysarthria.

This paper reports on measurements of several acoustic attributes of the fricative consonant /s/ produced in word-initial position by normally speaking adults and by speakers with neuromotor dysfunctions. Several acoustic properties are evaluated: the spectrum shape of the fricative and its amplitude in relation to the following vowel, the presence or absence of voicing, the time variation of the spectrum during the fricative and in the transition to the following vowel, and the presence of inappropriate acoustic patterns preceding the /s/. Some of these properties are based on quantitative measurements of the spectrum of the /s/, and others are based on observations of the time-varying acoustic patterns in spectrograms. For the individuals with dysarthria, deviations of each of these properties from the normal range are interpreted in terms of specific deficits in the control of the speech-production system. For the most part, these parameters are highly correlated with the speakers' overall intelligibility, with the intelligibility of words containing the fricative /s/, and with perceptual ratings of the adequacy of the fricative production. The parameters that show the best correlation with intelligibility and perceptual ratings are (a) measures of deviations from normalcy in the time variation of the acoustic pattern within the consonant and at the consonant-vowel boundary and (b) the spectrum shape of the frication noise. These acoustic parameters are related to deviations in the temporal pattern of control of the articulators in producing fricative-vowel sequences and to lack of fine control of the tongue blade in achieving an appropriate target configuration for the fricative.

Adult↗
PubMed2001

The role of phosphoinositide 3-kinase in the sorting and transport of newly synthesized tyrosinase-related protein-1 (TRP-1).

Tyrosinase-related protein-1 (TRP-1) is a 75 kDa type-1 transmembrane glycoprotein localized to the melanosome. The mechanism by which newly synthesized TRP-1 reaches its ultimate destination is currently unknown, but has been speculated to occur via the endosomal pathway. Recently, it has been shown that phosphatidylinositide (PI) 3-kinase is involved in various cellular functions, including regulating the constitutive movement of proteins from one intracellular compartment to another; however, whether PI 3-kinase participates in the trafficking of proteins such as TRP-1 to the melanosome is unknown. In this study we investigate the role of PI 3-kinase on the trafficking of TRP-1 in human melanoma MeWo cells using wortmannin, a potent inhibitor of PI 3-kinase. Our investigations demonstrate that wortmannin interferes with the membrane trafficking of TRP-1 in MeWo cells, and that it specifically results in the redistribution of the protein within a novel vesicular compartment with characteristics of the endosomal and lysosomal compartments [positive for LAMP-1, and partially positive for CD63 and cation-independent mannose 6-phosphate receptors (CI-M6PR)], and is accessible to internalized proteins such as immunoglobulins. Movement within this novel compartment is microtubule and GTPase dependent. These findings have led us to postulate that TRP-1 is sorted from the trans-Golgi network to a compartment in the vicinity of late endosomes, trafficking from which to the melanosome appears to be dependent on PI 3-kinase as it is blocked by wortmannin.

Androstadienes↗
PubMed2001

Increased sensitivity of melanocytes to oxidative stress and abnormal expression of tyrosinase-related protein in vitiligo.

BACKGROUND: Vitiligo is a depigmenting disease of the skin, which may derive from programmed melanocyte death or destruction due to inherent sensitivity to oxidative stress arising from either toxic intermediates of melanin, a melanocyte-specific protein, or other sources. Tyrosinase-related protein (TRP) -1 has been shown to be involved not only in melanin biosynthesis but also in the prevention of premature melanocyte death in animals. OBJECTIVES: To clarify the biological role of human TRP-1 in melanocyte survival. METHODS: Cultured melanocyte strains from an active advancing border of vitiligo were established and studied. RESULTS: The established 'vitiligo melanocytes' showed large perikaryon and stubby dendrites. They showed early cell death when exposed to oxidative stress (ultraviolet B) and increased and abnormal immunostaining and immunoprecipitation by antibodies against human and mouse TRP-1, indicating an altered synthesis and processing of TRP-1. In pulse-chase and sequential immunoprecipitation experiments, vitiligo melanocytes revealed abnormal protein-protein interaction with calnexin, a melanogenesis-associated chaperone, suggesting altered folding and maturation of nascent TRP-1 polypeptides. Northern blot analysis indicated a decreased expression of TRP-1 mRNA, but heteroduplex analysis and verification of the mutation at the carboxy terminus of TRP-1 by restriction enzyme analysis did not show any abnormality. CONCLUSIONS: Our study suggests that the early cell death of vitiligo melanocytes is related to their increased sensitivity to oxidative stress, which may arise from complex processes of abnormal synthesis and processing of TRP-1 and its interaction with calnexin.

Adult↗
PubMed2001

Modulation of M-channel conductance by adenosine 5' triphosphate in bullfrog sympathetic B-neurones.

1. Adenosine 5' triphosphate (ATP) (0.5-500 microM) or muscarine (0.1-1.0 microM) suppressed M-current/conductance (IM/gM) in B-cells of bullfrog sympathetic ganglion. Both agonists suppressed steady-state M-conductance (gM) at -30 mV and there was either no change or a slight increase in the time constants for gM activation (tau(a) at -30 mV) and deactivation (tau(d) at -50 mV). 2. It has previously been shown that experimental elevation of intracellular Ca2+ concentration ([Ca2+]i) suppresses gM and this is associated with decreases in both tau(a) and tau(d). As these changes in kinetics differ from those we observe with agonist application, our results cast doubt on the hypothesis that elevation of [Ca2+]i is involved in the transduction mechanism for ATP- or muscarine-induced gM suppression.

Adenosine Triphosphate↗
PubMed2001

IFN-gamma gene therapy by intrasplenic hepatocyte transplantation: a novel strategy for reversing hepatic fibrosis in Schistosoma japonicum-infected mice.

Liver-targeted gene therapy using hepatocyte as recipient cells has recently been documented to be effective in treatment of numerous hepatic diseases, such as metabolic diseases and liver carcinoma. IFN-gamma elicits antipreliferative and antifibrogenic activity in a variety of mesenchymal cells, including hepatic satellite cells. To investigate the antifibrogenic response of liver gene therapy mediated by intrasplenic transplantation of gene-modified hepatocytes, normal mouse liver cell line BNL CL.2 cells were transfected with murine IFN-gamma gene (BNL.IFN-gamma) in vitro, and transplanted intrasplenically into Schistosoma japonicum-infected mice. The amounts and distribution of IFN-gamma (which inhibits collagen synthesis), TGF-beta (which stimulates collagen synthesis) and extracellular matrix, including type I and III collagen, were detected. In mice infected with S. japonicum and then treated with BNL.IFN-gamma, an increase of IFN-gamma and decrease of TGF-beta1 were detected at 20 weeks postinfection compared to untreated S. japonicum-infected mice. Immunohistochemical analysis showed that S. japonicum infection induced a marked increase of type I and III collagen synthesis. Whereas, 4 weeks after treatment with BNL.IFN-gamma, net synthesis rates of type I and III collagen were markedly decreased in the liver of infected mice. In addition, a decreased expression of TGF-beta1 and its receptor TGF-betaRII in the liver of BNL.IFN-gamma-treated mice was also observed. Moreover, the decrease in TGF-beta1 and TGF-betaRII protein approximately paralleled the decrease in their mRNA expression, which was detected by RNA dot blotting. The data indicate that intrasplenic transplantation of IFN-gamma gene-modified hepatocyte can be a candidate approach to treat hepatic fibrosis.

Animals↗
PubMed2001

Learning needs of Taiwanese nurses caring for patients with irritable bowel syndrome.

This study, building on work on irritable bowel syndrome conducted by Letson and Dancey (1996), explores Taiwanese nurses' knowledge, perceptions and beliefs about caring for patients with irritable bowel syndrome (IBS). A 46-item questionnaire was completed voluntarily by 120 registered nurses from a large tertiary acute care facility in Taiwan. The study used a descriptive research design and a questionnaire was developed that used a five-point Likert scale (strongly agree = 5, to strongly disagree = 1). The questionnaire consisted of six categories: demographic information; nurses' sources of IBS information; nurses' knowledge about IBS; nurses' perceptions about patients with IBS; nurses' beliefs about IBS; and learning requirements for nurses. Overall, the results indicted that Taiwanese nurses who participated in this study had little specific knowledge of IBS. The researchers developed a booklet containing information on the condition of IBS that may be used by the participants in this study to fill the knowledge gaps about this condition and provide useful information.

Attitude of Health Personnel↗
PubMed2001

Histologic and immunophenotypic classification of cervical carcinomas by expression of the p53 homologue p63: a study of 250 cases.

Recent studies of the p53 homologue p63 indicate that this gene is preferentially expressed in basal and immature cervical squamous epithelium. This study correlated p63 expression with morphologic phenotype and human papillomavirus (HPV) type in a wide range of cervical neoplasms. Two hundred fifty cases of cervical carcinoma, including squamous cell carcinoma (SCCA; n = 178), adenocarcinoma (ADCA; n = 28), adenosquamous carcinoma (ASCA; n = 8), neuroendocrine carcinoma (NECA; n = 15), and other variant or mixed types (n = 21) were studied. Ninety-seven percent of SCCA, 0% of ADCA, and 0% of SCUC showed strong (>75% v <30%) positivity for p63 (P<.001). p63 sharply distinguished SCCA (p63+) from ADCA (p63-), Large-cell, poorly differentiated carcinomas were distinguished as putative glandular (glassy cell) or squamous (lymphoepithelial-like or spindle cell) types based on p63 staining. Eight (73%) of 11 neuroendocrine tumors tested were chromogranin positive; all showed no or low (<30%) levels of p63 immunostaining. Absence of p63 was also associated with a subset of nonneuroendocrine undifferentiated carcinomas. Transitions from squamous to columnar or undifferentiated morphology coincided with loss of p63 expression. A strong association between HPV 16 and p63 positivity was identified because of the colocalization of both within tumors of squamous phenotype. p63 is a powerful marker for squamous differentiation and, when diffusely expressed, excludes a glandular or neuroendocrine differentiation. p63 may be useful for differentiating pure squamous or glandular from adenosquamous carcinomas, tracking shifts in differentiation within tumors, supporting (by its absence) the diagnosis of neuroendocrine carcinomas, and clarifying the spectrum of poorly differentiated carcinomas lacking either squamous or neuroendocrine differentiation.

Adenocarcinoma↗
PubMed2001

Edge deepithelialization: a method to prevent leakage when tubed free skin flap is used for pharyngoesophageal reconstruction.

BACKGROUND: Anastomosis leakage remains a major source of morbidity and mortality in patients who receive pharyngoesophageal reconstruction, especially when a free skin flap is used as the substitute. This makes free skin flaps less favored than other substitutes in pharyngoesophageal reconstruction. However, a free skin flap provides many advantages for pharyngoesophageal reconstruction and, in some circumstances, may be the only choice. To reduce the incidence of postoperative leakage, we developed a technical modification called edge-deepithelialization for skin flap to ensure a more secure anastomosis. METHODS: Twenty-four patients who received tubed free skin flaps for cervical esophagus or hypopharynx reconstruction during a 7-year period were included in this study. Group 1 (n = 12) received flap edge-deepithelialization before undergoing the anastomosis, and Group 2 (n = 12) received the conventional anastomosis method. Preoperative nutritional status was evaluated by serum albumin level. All the patients were followed up for at least 1 year (1 approximately 14 years; mean, 7.3 years). Postoperative leakage was detected by clinical symptoms and signs or by routine esophagography. RESULTS: Postoperative anastomosis leakage developed in 1 patient in Group 1 (8.3%) and 5 patients in Group 2 (41.67%). The difference in the leakage rate between these groups was of statistical significance (P =.0001). There were no significant differences in the preoperative serum albumin level noted between these 2 groups (t test, P >.05). Older age was not associated with a higher leakage rate. Among the 6 leakage cases, 1 was treated conservatively, and the other 5 needed an additional 1 to 5 procedures for the management of leakage. All the leakages were cured during the follow-up period. CONCLUSIONS: With the flap edge deepithelialized, a more secure and reliable anastomosis can be obtained. This makes the free skin flap an easier and safer alternative in pharyngoesophageal reconstruction, especially when laparotomy and enteric flaps are contraindicated.

Adult↗
PubMed2001

Parathyroidectomy promotes wound healing and prolongs survival in patients with calciphylaxis from secondary hyperparathyroidism.

BACKGROUND: Calciphylaxis is a rare but life-threatening condition occasionally affecting patients with secondary hyperparathyroidism. Parathyroidectomy has been advocated as the only potentially curative intervention. METHODS: Between January 1989 and May 2000, 13 patients with pathologic/clinical criteria of calciphylaxis were treated at our institution. Of these 13 patients, 7 were managed with medical therapy alone, and 6 were referred for parathyroidectomy. The medical records were reviewed, and patients/relatives were interviewed. RESULTS: All patients had cutaneous wounds requiring local debridement predominantly located on the lower extremities or abdominal wall. Six patients underwent subtotal (3.5 gland) parathyroidectomy without morbidity. All 6 had significant reductions in parathyroid hormone levels after surgery (mean decrease, 94% +/- 0%), and all reported resolution of pain and healing of cutaneous wounds. Of the remaining 7 patients who had medical management alone, 5 eventually died of complications related to calciphylaxis. In comparing the 2 groups, patients who underwent parathyroidectomy had a significantly longer median survival than those who did not have surgery (36 vs 3 months, P =.021). CONCLUSIONS: Calciphylaxis frequently causes gangrene, sepsis, and eventual death. Parathyroidectomy can be performed with minimal morbidity and is associated with resolution of pain, wound healing, and a significantly longer median survival. Therefore, patients with secondary hyperparathyroidism and signs/symptoms of calciphylaxis should be referred promptly for consideration of parathyroidectomy.

Adult↗
PubMed2001

Removal of slowly biodegradable COD in combined thermophilic UASB and MBBR systems.

Starch, cellulose and polyvinyl alcohol (PVA) are common substrates of the slowly biodegradable COD (SBCOD) in industrial wastewaters. Removal of the individual and mixed SbCOD substrates was investigated in a combined system of thermophilic upflow anaerobic sludge blanket (TUASB) reactor (55 degrees C) and aerobic moving bed biofilm reactor (MBBR). The removal mechanisms of the three SBCOD substrates were quite different. Starch-COD was almost equally utilized and removed in the two reactors. Cellulose-COD was completely (97-98%) removed from water in the TUASB reactor by microbial entrapment and sedimentation of the cellulose fibers. PVA alone was hardly biodegraded and removed by the combined reactors. However, PVA-COD could be removed to some extent in a binary solution of starch (77%) plus PVA (23%). The PVA macromolecules in the binary solution actually affected the microbial activity in the TUASB reactor resulting accumulation of volatile fatty acids, which shifted the overall COD removal from the TUASB to the MBBR reactor where SBCOD including PVA-COD was removed. Since the three SBCOD substrates were removed by different mechanisms, the combined reactors showed a better and more stable performance than individual reactors.

Bacteria, Anaerobic↗