PubMed Health⌕ Search

Biomedical subjects

C L Ho

Publications and source records attributed to C L Ho.

At least 55 records · Page 3Linked to original sources
PubMed1998

Structures and biological activities of new wasp venom peptides isolated from the black-bellied hornet (Vespa basalis) venom.

The black-bellied hornet (Vespa basalis) is the most dangerous species of vespine wasps found in Taiwan. The hornet possesses a highly toxic venom which is rich in toxin, enzymes and biologically active peptides. Using ultrafiltration to remove the high molecular weight toxin and enzymes followed by reverse-phase HPLC, three bioactive tridecapeptides, designated "HP-1, HP-2 and HP-3" were isolated from the venom. Their amino acid sequences were determined as: HP-1: LFRLIAKTLGSLM, HP-2: LFRLLANTLGKIL, HP-3: IFGLLAKTLGNLF. The primary structures of these peptides appear to be homologous to those of chemotactic peptides isolated from other vespid venoms. However, these peptides show little chemotactic activity on human neutrophils and have distinct tripeptide sequences at the amino and carboxylic terminal sides, as compared with other hornet chemotactic peptides. The lack of Pro3 which is a characteristic structure of vespid chemotactic peptides in their sequences is most distinctive. Circular dichroism spectra of these peptides measured in 20% trifluoroethanol show a high content of alpha-helical conformation. All three peptides provoked local edema in rat hind paw, which could be inhibited by antihistamine (diphenhydramine) and drug with antiserotonin activity (cyproheptadine). These peptides also exhibited a potent hemolytic activity which was potentiated by a non-lytic dose of the hornet lethal protein, suggesting a supporting role of these peptides in the lethal effect of Vespa basalis venom.

Amino Acid Sequence↗
PubMed1998

Molecular characterization of plastidic phosphoserine aminotransferase in serine biosynthesis from Arabidopsis.

Serine biosynthesis in plants proceeds by two pathways; a photorespiratory pathway which is associated with photorespiration and a pathway from phosphoglycerate. A cDNA encoding plastidic phosphoserine aminotransferase (PSAT) which catalyzes the formation of phosphoserine from phosphohydroxypyruvate has been isolated from Arabidopsis thaliana. Genomic DNA blot analysis indicated that this enzyme is most probably encoded by a single gene and is mapped on the lower arm of chromosome 4. The deduced protein contains an N-terminal extension exhibiting the general features of a plastidic transit peptide, which was confirmed by subcellular organelle localization using GFP (green flourescence protein). Northern analysis indicated preferential expression of PSAT in roots of light-grown plants, supporting the idea that the phosphorylated pathway may play an important role in supplying the serine requirement of plants in non-green tissues. In situ hybridization analysis of PSAT revealed that the gene is generally expressed in all types of cells with a significantly higher amount in the meristem tissue of root tips.

Amino Acid Sequence↗
PubMed1998

Peripherin is tyrosine-phosphorylated at its carboxyl-terminal tyrosine.

Peripherin is a type III intermediate filament present in peripheral and certain CNS neurons. We report here that peripherin contains a phosphotyrosine residue and, as such, is the only identified intermediate filament protein known to be modified in this manner. Antiserum specific for phosphotyrosine recognizes peripherin present in PC12 cells (with or without nerve growth factor treatment) and in rat sciatic nerve as well as that expressed in Sf-9 cells and SW-13 cl. 2 vim- cells. The identity of peripherin as a tyrosine-phosphorylated protein in PC12 cells was confirmed by immunoprecipitation, two-dimensional isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels, and phosphoamino acid analysis. Unlike serine/threonine phosphorylation, tyrosine phosphorylation of peripherin is not regulated by depolarization or nerve growth factor treatment. To identify the site of tyrosine phosphorylation, rat peripherin was mutated at several tyrosine residues and expressed in SW-13 cl. 2 vim- cells. Tyrosine phosphorylation was selectively lost only for peripherin mutants in which the carboxy-terminal tyrosine (Y474) was mutated. Indirect immunofluorescence staining indicated that both wild-type peripherin and peripherin Y474F form a filamentous network in SW-13 cl. 2 vim- cells. This indicates that tyrosine phosphorylation of the peripherin C-terminal residue is not required for assembly and leaves open the possibility that this modification serves other functions.

Amino Acid Sequence↗
PubMed1998

Ultrastructural studies of carboxyl-terminal truncation mutants of the neuronal intermediate filament protein peripherin.

We have prepared carboxyl-terminal truncation mutants of the neuronal intermediate filament (IF) protein peripherin and examined the assembly characteristics of these mutant proteins in SW13 cells in the presence and absence of vimentin. In the absence of vimentin, tailless peripherin protein (Per-C424) self-assembles into bundles and clumps as observed by immunofluorescence, whereas a peripherin mutant that is missing the tail as well as a small portion of the rod (Per-C356) appears as spherical aggregates. Similar phenotypes are observed when vimentin-positive cells are transfected with Per-C424 or Per-C356. In these cells, the entire IF network is disrupted, and vimentin colocalizes with the mutant peripherin proteins. To examine the morphology of the bundles and clumps formed by Per-C424 at the electron microscopic level, we prepared stable cell lines expressing different levels of this mutant protein. By immunofluorescence, Per-C424 appears as either clumps or bundles of filaments depending on the expression level of the mutant protein. However, under electron microscopy, it is apparent that both clumps and bundles are composed of tightly packed IFs. We were unable to obtain stable cell lines expressing Per-C356, indicating that this mutant may prevent cell proliferation. Using a vector containing an internal ribosomal entry site, we prepared a construct that expresses Per-C356 and green fluorescent protein as a single mRNA, and we were able to isolate cells that expressed Per-C356 by fluorescence-activated cell sorting. Electron microscopic analysis of these cells showed that these aggregates are solid and contain no obvious filamentous structures.

Cell Line↗
PubMed1998

Awareness of environmental issues and the acceptance of CFC-free inhalers.

With the recent availability of a chlorofluorocarbon (CFC)-free metered dose inhaler (MDI) (Airomir), a patient survey was carried out to evaluate awareness of the role of CFCs in our environment and acceptance of this new inhaler. A questionnaire survey was conducted on parents and guardians of 201 children. Depending on respondents' preference, the interview was conducted in English (71%), Chinese (23%), Malay (5%) or Tamil (1%). A 'taste' test was also conducted on 103 of these children. Only 13% (26/201) of parents/guardians were aware that MDIs contained CFCs. Although 70% of children were in favour of the new taste of the CFC-free inhaler, the cost of the new inhaler was an important consideration for parents and guardians in their decision to switch to the new inhaler. The majority (93%) were willing to switch if its cost were equivalent to their current inhaler. This study has provided pertinent information with regard to acceptance of CFC-free inhalers which should be considered when making the inevitable switch to environmentally friendly inhalers.

Adult↗
PubMed1998

Novel features of intermediate filament dynamics revealed by green fluorescent protein chimeras.

In order to study the dynamic behavior of intermediate filament networks in living cells, we have prepared constructs fusing green fluorescent protein to intermediate filament proteins. Vimentin fused to green fluorescent protein labeled the endogenous intermediate filament network. We generated stable SW13 and NIH3T3 cell lines that express an enhanced green fluorescent protein fused to the N-terminus of full-length vimentin. We were able to observe the dynamic behavior of the intermediate filament network in these cells for periods as long as 4 hours (images acquired every 2 minutes). In both cell lines, the vimentin network constantly moves in a wavy manner. In the NIH3T3 cells, we observed extension of individual vimentin filaments at the edge of the cell. This movement is dependent on microtubules, since the addition of nocodazole stopped the extension of the intermediate filaments. Injection of anti-IFA causes the redistribution or 'collapse' of intermediate filaments. We injected anti-IFA antibodies into NIH3T3 cells stably expressing green fluorescent protein fused to vimentin and found that individual intermediate filaments move slowly towards the perinuclear area without obvious disassembly. These results demonstrate that individual intermediate filaments are translocated during the collapse, rather than undergoing disassembly-induced redistribution. Injections of tubulin antibodies disrupt the interactions between intermediate filaments and stable microtubules and cause the collapse of the vimentin network showing that these interactions play an important role in keeping the intermediate filament network extended. The nocodazole inhibition of intermediate filament extension and the anti-IFA microinjection experiments are consistent with a model in which intermediate filaments exhibit an extended distribution when tethered to microtubules, but are translocated to the perinuclear area when these connections are severed.

3T3 Cells↗
PubMed1998

Acute retinal necrosis--early manifestation and successful treatment with steroid and acyclovir: case report.

A healthy 19-year-old woman suffering from pain, redness, and blurred vision of her left eye came to our emergency unit for help. Initial examination revealed no light-perception, tenderness and marked inflammation of the left eye. There was severe inflammation both in the anterior chamber and vitreous cavity. Total retinal detachment accompanied by diffuse thickening of choroid was demonstrated using echography. Under the impression of panuveitis, oral steroids were given. The response of the left eye was dramatic but the patient complained of blurred vision of her right eye 2 days after steroid therapy. There was only mild reaction in the right eye; however, multiple granule-appearing white dots were found on the inferior two-thirds of the peripheral retina. The nummular white lesions increased in size and number and finally became confluent in the following days. The diagnosis was shifted to acute retinal necrosis and intravenous acyclovir was started while continuing systemic steroid therapy. The retinal necrosis began to consolidate four days after the administration of acyclovir and gradually regressed. We tapered the systemic steroids when the intra-venous acyclovir was shifted to oral form. The vitreous of the right eye remained clear during the follow-up period. The antiviral agent still suppressed the disease process even though steroids had been used beforehand. Furthermore, the systemic steroids seemed to ameliorate the vitritis and secondary complications in the right eye compared with the fulminant, disastrous course of the left eye.

Acyclovir↗
PubMed1997

The role of the N-terminal leucine residue in snake venom cardiotoxin II (Naja naja atra).

The N-terminal leucine residue of snake venom cardiotoxin II (CTX II) (Naja naja atra) was systematically replaced with D-leucine (CTXII-L1-D-L), glycine (CTXII-L1G) or deleted [CTXII-(2-60)] to study the role of leucine residue in CTX II molecule. CTX II, CTXL1-D-L, CTXL1G and CTX(2-60) were produced by chemical synthesis method and purified by high performance liquid chromatography. Owing to folding problem in CTXII-(2-60), only CTX II, CTXII-L1-D-L and CTXII-L1G were produced in a pure form and characterized by amino acid analysis, mass spectrometry and peptide mapping. In the structural aspect, changing the Leu-1 by D-Leu or Gly causes a drastic alteration in the whole CTX II structure as detected by circular dichroism, 1-anilino-naphthalene-8-sulfonate (ANS) fluorescence assay. In the functional aspect, both CTXII-L1-D-L and CTXII-L1G are still retained substantial biological activity of CTX II. Therefore, the results indicate that both the chirality and the side-chain of the N-terminal leucine residue of CTX II are important elements in maintaining the whole CTX II structure. In addition, this study is the first report in elucidating the reason why the first N-terminal residue of most CTXs (90.3%) is leucine residue.

Amino Acids↗
PubMed1997

Intraocular injection of recombinant hirudin to prevent experimental postoperative fibrin.

PURPOSE: To test the efficacy of a single intraocular injection of various doses of recombinant desulphatohirudin variant 1 (Revasc) in a rabbit model after lensectomy-vitrectomy surgery. METHODS: Standard fragmatome lensectomies and core vitrectomies were performed on Dutch Belted rabbits. In a masked fashion, 17 control eyes received single intraocular injections of lactated Ringer's solution, and the treated eyes, with eight eyes per treatment group, received single intraocular injections of hirudin at doses of 0.02 microgram, 2 micrograms, 10 micrograms, 50 micrograms, or 200 micrograms. RESULTS: Intraocular injections of hirudin were effective in preventing postoperative fibrin formation at doses of 2 micrograms. There was no notable intraoperative bleeding. With an injection of 2.0 micrograms hirudin there was no hemorrhage at any time after surgery; with an injection of 200 micrograms hirudin, however, there was notable bleeding. CONCLUSION: Hirudin is effective in the prevention of postoperative fibrin formation with a single intraocular injection of 2.0 micrograms after lensectomy-vitrectomy surgery in the rabbit model. There was no bleeding at the effective dose of 2.0 micrograms; at 100 times the effective dose, however, there was significant postoperative bleeding.

Animals↗
PubMed1997

Infections in paediatric renal transplant recipients.

A retrospective study of all children who had undergone renal transplantation at the National University Hospital was undertaken in order to investigate the types and frequency of infections in paediatric renal transplant recipients. A total of 13 children ranging in age from 3 to 17 years were transplanted over a 7-year period from 1989 to 1996. The mean follow-up period was 40.5 months. Ten patients (77%) had at least one episode of infection post-transplant, 70% of which occurred within the first 6 months post-transplant. These infections included urinary infections (8), disseminated cytomegalovirus (CMV) disease (4), Pneumocystis carinii pneumonia (1) and mycoplasma pneumonia (1). Urinary tract infections were significantly associated with the presence of ureteral stents. CMV disease was significantly associated with the use of mycophenolic acid in the early post-transplant period. Infection therefore continues to be a major cause of morbidity in paediatric renal transplant recipients.

Adolescent↗
PubMed1996

Differential expression of adenosine A1 receptors in colorectal cancer and related mucosa.

Adenosine A1 receptors (A1R) are known to inhibit while the A2 receptors (A2R) stimulate the G-protein cAMP second messenger system and may play a role in cell growth and carcinogenesis. Using a quantitative reverse transcription-polymerase chain reaction (RT-PCR) method, mRNA for A1R and A2R was measured in human colorectal adenocarcinomas and individual peritumoural colon tissues. There was differential expression of the mRNA for A1R with tumour tissues having significantly higher amounts compared to peritumoural normal tissues. The mRNA for A2R was not found to be differentially expressed. The increase in the inhibitory A1 receptor in tumour tissues may be in response to increased adenosine release from the hypoxic cells found in malignant tumour tissues, thus indicating a possible role for the adenosine A1 receptor in carcinogenesis.

Adult↗
PubMed1996

Intermediate filaments in the nervous system: implications in cancer.

In this review, we describe the different intermediate filament (IF) proteins, their assembly into IFs, the functions of IFs and their relation to disease with a particular emphasis on the intermediate filaments expressed in the nervous system. In the mammalian nervous system, seven intermediate filament proteins are known to be expressed in neurons or neuroblasts. These include the three neurofilament triplet proteins, which are present in both central and peripheral neurons; alpha-internexin, which is the first neuronal intermediate filament protein expressed in the developing mammalian nervous system and present primarily in CNS neurons in the adult nervous system; peripherin, which is most abundant in the PNS; vimentin, which is expressed in neuronal progenitor cells along with nestin, as well as in a few adult neurons. In contrast to these neuron-specific IF proteins, the glial fibrillary acidic protein (GFAP) is glial specific and expressed in mature astrocytes. Vimentin and nestin are also expressed in glial progenitor cells and vimentin is expressed along with GFAP in some mature astrocytes. As a whole, the expression of IF proteins is tissue specific and developmentally regulated. As a result, IF proteins are good markers for determining the cell origin and differentiation status of tumor cells. For example, peripherin is expressed in neuroblastomas, GFAP in astrocytomas and neurofilaments in tumors of neuronal origin. However, tumor cells may express IF patterns which are irrelevant to their cell origin. Therefore, one has to be very careful in using IF patterns as sole indicators of cell origin and differentiation status of tumors.

Animals↗
PubMed1996

Structural requirements for the edema-inducing and hemolytic activities of mastoparan B isolated from the hornet (Vespa basalis) venom.

Mastoparan B (MP-B) is a cationic tetradecapeptide isolated from the black-bellied hornet (Vespa basalis) venom. It has a primary structure (LKLKSIVSWAKKVL-CONH2) distinct from other vespine mastoparans. The peptide caused a dose-dependent swelling in rat hind paw and showed a potent hemolytic activity in guinea pig red blood cells. Studies on the structure activity relationship of the peptide showed that replacing lysine at position 2 (Lys2) by asparagine (Asn) in the MP-B sequence caused about 40% decrease in its edema-inducing activity at 50 micrograms/paw and 90% decrease in hemolytic activity at 30 microM of the peptide, while the same substitution at Lys4 did not cause a significant change in either activity. Replacing either Lys11 or Lys12 by leucine (Leu) caused little or no decrease in the edema-inducing and hemolytic activities. Decreases in both activities were observed when both Lys11 and Lys12 were replaced by Leu. On the other hand, replacing tryptophan at position 9 (Trp9) by tyrosine or phenylalanine in MP-B sequence almost abolished its hemolytic activity, while the edema-inducing activity was only partially inhibited. Circular dichroism spectra of the peptides measured in 20% trifluoro-ethanol revealed that substitution of Lys and Trp did not cause a significant change in the conformation of MP-B. it appears that Lys2 is crucial for both hemolytic and edema-inducing activities of MP-B, while Trp9 is of special importance to the hemolytic activity of MP-B. Lys11 and Lys12 in MP-B probably play a lesser role in both activities.

Amino Acid Sequence↗
PubMed1996

[Myxoma of the left atrium causing acute myocardial infarction].

A 66-year-old male patient had an acute inferior wall myocardial infarction 55 days prior to the admission to the hospital. Echocardiogram and cardiac catheterization disclosed a tumor of the left atrium and the coronary arteriography was normal. The tumor was surgically removed and pathological examination revealed it to be a myxoma. In the absence of other etiologic possibilities, left atrial myxoma was considered the main hypothesis with the development of thrombi and coronary embolization.

Heart Atria↗
PubMed1995

Molecular cloning and characterization of a neurotoxic phospholipase A2 from the venom of Taiwan habu (Trimeresurus mucrosquamatus).

Using gel-filtration chromatography and reverse-phase (RP) HPLC we have purified a presynaptic neurotoxin (designated as trimucrotoxin) from the crude venom of Taiwan habu (Trimeresurus mucrosquamatus). Its complete primary structure was solved by an automated N-terminal sequencing and cDNA sequencing method. The enzyme inhibited the twitch of the chick biventer cervicis muscle at 0.1-1 micrograms/ml and showed lethality in mice (LD50 = 1.2 micrograms/g, when given intravenously). Trimucrotoxin exists mainly as a homodimer of 14 kDa subunits as shown by a gel-filtration experiment, and dissociates into monomers during SDS/PAGE in the absence of Ca2+. However, most of trimucrotoxin migrated as slowly as a trimer during nondenaturing SDS/PAGE in the presence of Ca2+ or Sr2+. Its amino acid sequence identity to crotoxin B and agkistrodotoxin is about 75%, and its cDNA sequence is 82% identical to that of crotoxin B. Rabbit antiserum against trimucrotoxin also cross-reacted with the other crotalid neurotoxic phospholipases A2. Furthermore, the purified acidic subunit of crotoxin potentiated the neurotoxicity of trimucrotoxin. A comparison of the sequences of these crotalid neurotoxins revealed some common features of the possible neurotoxic sites, including residues 6, 11, 76-81 and 119-125.

Amino Acid Sequence↗