Biomedical subjects

J S Sun

Publications and source records attributed to J S Sun.

At least 37 records · Page 2Linked to original sources

PubMed2000

Recognition and cleavage of DNA by rebeccamycin- or benzopyridoquinoxaline conjugated of triple helix-forming oligonucleotides.

Indolocarbazole and benzopyridoquinoxaline derivatives have been shown to have anti-tumor activity and to stimulate DNA topoisomerase I-mediated cleavage. Two indolocarbazole compounds (R-6 and R-95) and one benzopyridoquinoxaline derivative (BPQ(1256)) were covalently attached to the 3'-end of a 16mer triple helix-forming oligonucleotide (TFO). These conjugates bind to DNA with a higher affinity than the unsubstituted oligonucleotides. Furthermore, they induce topoisomerase I-mediated and triplex-directed DNA cleavage in a sequence-specific manner.

Aminoglycosides↗

PubMed2000

Linkage of a triple helix-forming oligonucleotide to amsacrine-4-carboxamide derivatives modulates the sequence-selectivity of topoisomerase II-mediated DNA cleavage.

Amsacrine-4-carboxamide-oligonucleotide conjugates were synthesized and studied for their capacity to form DNA triple helices and to alter human topoisomerase II binding and cleavage properties. The intercalating agent was attached to the 3'- or the 5'-end of a 24 nt triple helix-forming oligonucleotide via linkers of different lengths. The stability of these DNA triple helices was investigated by gel retardation and melting temperature studies using a synthetic 70 bp DNA duplex target. The effect of the conjugates on DNA cleavage by topoisomerase II was evaluated using the 70 bp duplex and a 311 bp restriction fragment containing the same triple helix site. The conjugate with the amsacrine derivative linked to the 3' end of the TFO via a hexaethylene glycol linker modulates the extent of DNA cleavage by topoisomerase II at specific sites.

Amsacrine↗

PubMed2000

Mycobacterial infection of the upper extremities.

BACKGROUND AND PURPOSE: Mycobacterial infection of the upper extremities is extremely rare. To make an early accurate diagnosis is often difficult, and the treatment is, therefore, frequently delayed in clinical practice. This paper describes the diagnosis and treatment of mycoabacterial infection of the upper extremities in a series of patients treated at National Taiwan University Hospital. METHODS: The medical records of 15 patients with mycobacterial infection of the upper extremities treated between 1985 and 1998 were retrospectively analyzed. The diagnosis of mycobacterial infection was established by positive culture results, the presence of acid-fast bacilli, or characteristic histopathologic findings. RESULTS: Mycobacterial infection was suspected on initial examination in only two patients. The duration between the onset of symptoms and correct diagnosis averaged 37.5 months (range, 3 wk to 209 mo). The involved sites of mycobacterial infection included the wrist in nine patients, the elbow in two, the hand in two, the humerus in two, the forearm in one, and the shoulder in one. Three patients suffered from multifocal musculoskeletal mycobacterial infection. Microbiologic studies identified Mycobacterium tuberculosis in four patients and nontuberculous mycobacteria (NTM) in four. Characteristic histopathologic findings of mycobacterial infection including caseation, granulomatous inflammation, eosinophilic epithelioid cells, Langhan's giant cells, and lymphocytic aggregates were noted in all 15 patients. All patients received chemotherapy and 14 patients underwent surgical treatment. The mean duration of follow-up was 46 months (range, 15-97 months). Treatment failed in two of the patients with multifocal NTM infection and in one patient whose mycobacterial infection was identified by histopathology. CONCLUSIONS: Mycobacterial infection should be included in the differential diagnosis of infection of the upper extremities. Treatment failures are more frequent in patients with multifocal NTM infection.

Adolescent↗

PubMed2000

Ipsilateral distal radius and scaphoid fractures.

Simultaneous fractures of the distal radius and scaphoid are uncommon. Previous reports have outlined a variety of treatment methods for this condition. We describe the results in eight patients with simultaneous fractures of the distal radius and scaphoid, who were treated at National Taiwan University Hospital from 1987 to 1998. Treatments for the distal radius fractures included pin-in-cast in one patient, casting in one, external fixation in one, external fixation after pinning in three, and open reduction with internal fixation in two. All scaphoid fractures occurred at the scaphoid waist, and were treated with long-arm thumb spica in three patients, Herbert screw fixation in two, and percutaneous pinning in three. All of the fractures united, and none of the patients developed scaphoid avascular necrosis. One patient suffered from superficial radial nerve injury during external fixation. The functional results of the wrist were good in five patients and fair in three. To prevent the possible over-distraction of the scaphoid fracture, fixation of the scaphoid should be performed if a distraction force is to be applied during the treatment of the distal radius fracture. If no distraction force is to be applied, a thumb spica may provide adequate fixation for the scaphoid fracture.

Adult↗

PubMed1999

Optimization of alternate-strand triple helix formation at the 5"-TpA-3" and 5"-ApT-3" junctions.

Alternate-strand triple helix formation was optimized at the two junction steps, the 5"-TpA-3" and 5"-ApT-3" junctions. Footprint experiments, gel retardation assays and thermal denaturation measures on a sequence appropriately designed with two adjacent alternate-strand polypurine tracts points out that the addition of an adenine residue and the removal of one nucleotide should facilitate the crossing strands at the 5"-TpA-3" junction and at the 5"-ApT-3" junction, respectively. These results provide a 'switch code' for the construction of alternate-strand triple helix forming oligonucleotides which open new possibilities for extending the range of applications of antigene strategy.

Adenine↗

PubMed1999

The influence of hydroxyapatite particles on osteoclast cell activities.

Aseptic loosening after total joint arthroplasty is a major problem in orthopedic surgery. Small particles from material wear have been reported as the main cause of implant failure. For this reason, investigation into possible wear particles from the materials used in the implant may lead to longevity after arthroplasty. Hydroxyapatite (HA) has been extensively investigated and reported as an excellent biomaterial with excellent biocompatibility. In this study, we used an in vitro osteoblast/osteoclast model to test the biocompatibility of various-sized HA particles. Primary osteoclasts/osteoblasts were co-cultured with different-sized HA particles (0.5-3.0 microm, 37-53 microm, 177-205 microm, and 420-841 microm) for 3 h, 1 day, 3 days, and 7 days. Cellular responses to the HA particles were evaluated by changes in cell counts and the secretion of transforming growth factor (TGF-beta1), alkaline phosphatase (ALP), tumor necrosis factor (TNF-alpha), prostaglandin (PGE2), and lactate dehydrogenase (LDH) in the supernatant of the culture media. The results showed that osteoblasts/osteoclasts co-cultured with HA particles smaller than 53 microm undergo the most significant changes. Cellular counts significantly decreased, and the changes were more obvious in the osteoblast population. There also was a significant decrease in TGF-beta1 concentration and a significant increase in PGE2 and LDH concentration, but there were no changes in the TNF-alpha or ALP titer. It can be concluded that larger HA particles may be quite compatible with bone cells while smaller-sized HA particles can both activate the osteoclasts and decrease the cell population of the osteoblasts. Justification for the additional expense incurred with the use of hydroxyapatite in primary total hip arthroplasty should be further evaluated.

Alkaline Phosphatase↗

PubMed1999

Bone defect healing enhanced by ultrasound stimulation: an in vitro tissue culture model.

Ultrasound has many medical applications. Previous animal and clinical studies have clearly shown a positive effect of ultrasound on the rate of osseous repair. The present in vitro study was designed to elucidate the specific response of bony tissue to ultrasound treatment. Bilateral femora were obtained from 36 mature male Wistar rats. A bone defect was created at the center of each distal metaphysis. The femora were maintained for either 7 or 14 days in in vitro tissue culture and received 15 min of ultrasound stimulation or a sham exposure. The ultrasound intensity used was either 320 or 770 mW/cm2. Healing of the bone defect was evaluated by histomorphological examination and by analysis for the synthesis and secretion of prostaglandin E2. The results showed that ultrasound stimulation can accelerate both defect healing and trabecular bone regeneration. All experimental femoral defects treated with ultrasound healed faster than the untreated cortical defects, but only the defects receiving 770 mW/cm2 reached a level that was significantly different. The healing rate for the 320-mW/cm2 stimulated defects was intermediate between that of the 770-mW/cm2 and sham-exposed defects. With ultrasound stimulation, prostaglandin E2 secretion by the experimental femora decreased significantly. Changes in the prostaglandin synthesis and concentration were found to correspond to changes in the amount of trabecular regeneration and to acceleration of bone healing. This highly controlled and well-studied model of ultrasound stimulation of bone healing in vitro can be used to further examine the biological mechanisms involved.

Animals↗

PubMed1999

Preparation of a biphasic porous bioceramic by heating bovine cancellous bone with Na4P2O7.10H2O addition.

Sintered bovine cancellous bone exhibited excellent biocompatiblity, high porosity and have an interconnecting porous structure allowing for bone ingrowth. However, the main mineral constitution of sintered bovine bone-hydroxyapatite (Ca10(PO4)6(OH)2, HAP) seems to be too stable in vivo. For improving its bioactivity, the calcined bovine bone removing the organic substance by burning process-with different quantities of sodium pyrophosphate (Na4P2O7.10H2O, NP) addition was heated to a high temperature to transform its crystalline phase constitution from HAP into TCP/HAP biphasic or other multiphasic structures. Results revealed that the calcined bovine bone without NP addition, exhibited a pure form of HAP characterized pattern during heating. Its thermal behavior was similar to stoichiometric HAP, it gradually lost its OH- ions and transformed into oxyhydroxyapatite at high temperature. After being doped into calcined bovine bone, NP would react with HAP to form betaBTCP and NaCaPO4 around 600 degrees C. At 900 degrees C, doped NP would completely react with HAP and the NaCaPO4 would further react with HAP to form more betaBTCP in the system. With NP increasing in the calcined bovine bone, HAP would gradually convert into different crystalline phase compositions of TCP/HAP, TCP/HAP/NaCaPO4 or TCP/NaCaPO4 at high temperature. By heating calcined bovine cancellouse bone with different quantities of NP we could obtain different crystalline phase compositions of natural porous bioceramic in this study.

Animals↗

PubMed1999

Thermal decomposition and reconstitution of hydroxyapatite in air atmosphere.

In this paper, the decomposition and reconstruction behavior of hydroxyapatite (HAP) during heating and cooling in air atmosphere were studied. The commercial HAP were chosen and gradually heated to 1500 degrees C and cooled to room temperature by a program controlled SiC heated furnace. X-ray diffraction (XRD) and Fourier-transformed infrared (FTIR) analysis were used to investigate the change of crystalline phases and functional groups of HAP at different temperatures. Weight change of samples was recorded by thermogravimetric analysis (TGA) during heating and cooling. The results revealed that HAP gradually releases its OH- ions and transforms into OHAP in the temperature of 1000-1360 degrees C. Above 1360 degrees C, the OHAP would decompose into TTCP and alpha TCP phase. The OH- stretching bands of HAP could be traced by FTIR even at the temperature of 1350 degrees C which indicates HAP decomposition. HAP does not dehydrate completely before decomposition. We speculated that some oxyapatite (OAP) might be formed during dehydration with a great amount of OHAP still left in the system even up to the temperature of decomposition. In the temperature range of 1400-1500 degrees C, there was no significant difference in XRD patterns, only TTCP and alpha TCP crystalline phases were observed. When the HAP gradually cools from 1500 degrees C, a part of TTCP and alpha TCP would directly reconstruct into OAP around 1350 degrees C. OAP existed in the temperature range of 1350-1300 degrees C during cooling. When the temperature decreased to 1290 degrees C, a part of TTCP and alpha TCP reconstructed into OHAP by rehydration reaction and OAP were rehydrated into OHAP as well. At 1100 degrees C, the rest of TTCP and alpha TCP reconstitutes into HAP. As the temperature decreases, the OHAP is gradually rehydrated and reconstituted into HAP.

Air↗

PubMed1999

Targeting topoisomerase I cleavage to specific sequences of DNA by triple helix-forming oligonucleotide conjugates. A comparison between a rebeccamycin derivative and camptothecin.

Topoisomerase I is an ubiquitous DNA cleaving enzyme and an important therapeutic target in cancer chemotherapy for the camptothecins as well as for indolocarbazole antibiotics such as rebeccamycin and its synthetic derivatives, which stabilize the cleaved DNA-topoisomerase I complex. The covalent linkage of a triple helixforming oligonucleotide to camptothecin or to the indolocarbazole derivative R-6 directs DNA cleavage by topoisomerase I to specific sequences. Sequence-specific recognition of DNA is achieved by the triple helix-forming oligonucleotide, which binds to the major groove of double-helical DNA and positions the drug at a specific site. The efficacy of topoisomerase I-induced DNA cleavage mediated by the rebeccamycin-conjugate and the camptothecin-conjugate was compared and related to the intrinsic potency of the isolated drugs.

Aminoglycosides↗

PubMed1999

Scavenging effect of benzophenones on the oxidative stress of skeletal muscle cells.

Benzophenone is an ultraviolet (UV)-absorbing agent that has been used in industry and medicine for more than 30 years. Consumers of cosmetics and sunscreens containing UV-absorbers are exposed to benzophenones on a daily basis, owing to the widespread use of these compounds. However, the efficacy of these compounds as scavengers of oxidative stress is still not well established. In the present study, we investigate the antioxidative capacity of six sunscreen benzophenone compounds. A primary myoblast culture was mixed in vitro with 100 microM menadione. The cytotoxic effect by menadione-induced oxidative stress was monitored by the lucigenin- or luminol-amplified chemiluminescence, methylthiotetrazole (MTT) assay, and the antioxidative effects of various benzophenone compounds were evaluated. The results showed that the addition of menadione can induce oxidative stress on myoblasts by superoxide and hydrogen peroxide production, which can be eradicated by superoxide dismutase (SOD) and catalase, respectively, in a dose-dependent mode. The catalase has a protective effect on the cytotoxicity induced by menadione as measured by the MTT assay, while the SOD does not. The selected benzophenones also have a significant scavenging effect on the menadione-induced cell death on the myoblasts. The ortho-dihydroxyl structure and other hydroxy groups in the same ring have a stronger scavenging effect on the superoxide anion on myoblasts; thus, a stable penoxy radical may be formed. The mechanism of this effect remains to be clarified.

Acridines↗

PubMed1999

The effect of sintered beta-dicalcium pyrophosphate particle size on newborn Wistar rat osteoblasts.

During recent years, sintered dicalcium phosphate (SDCP) has been shown to be an effective artificial bone filler for repairing bone defects. The goal of this study was to elucidate the effect of SDCP particle size on osteoblasts. Osteoblasts were mixed and cultured with various sized SDCP particles (0.5-3.0, 37-63, 177-250, and 420-841 microns) for 1 h, 3 h, 1 day, 3 days, and 7 days and then analyzed. The results show that the adding of smaller sized SDCP particles (0.5-3.0 and 37-63 microns) into osteoblast culture can significantly affect the cell counts of osteoblasts. The secretion of transforming growth factor-beta 1, alkaline phosphatase, and prostaglandin E2 in culture medium increased significantly. The changes were most significant and persisted longer in smaller particle groups. Small sintered dicalcium phosphate particles can inhibit the proliferation of the osteoblasts. The inhibitory effects of the smaller sized SDCP particles on the osteoblasts were mediated by the promotion of osteoblast differentiation and the increased synthesis of prostaglandin E2.

Alkaline Phosphatase↗

PubMed1999

Antioxidant status following acute ischemic limb injury: a rabbit model.

Although ischemic injury to skeletal muscle is a matter of great clinical importance, relatively little is known about the mechanisms which determine systemic responses. One purpose of this study is to elucidate the systemic antioxidant status following an episode of acute ischemic limb injury and subsequent reperfusion. Twelve New Zealand white rabbits were used in this study. After the animals were anesthetized, an ischemic insult was created in the right hind limb for twelve hours, followed by four hours of reperfusion. Several series of blood samples were obtained. At the end of the experiment, the animals were killed and necropsies undertaken in order to evaluate the antioxidant status of various visceral organs. The results link ischemia and reperfusion injury to a significant decline in antioxidative activity in various tissues. The weakening in antioxidant status after ischemic limb injury was most pronounced in the heart tissue, followed in descending order by the spleen, skeletal muscle, lung, liver, and kidney tissue. The levels of specific antioxidants and reactive oxygen species in various organs changed significantly, and the changes were tissue specific. Endogenous radical scavenging systems were not entirely overwhelmed in most of the tissues studied. But higher levels of malondialdehyde (MDA) found in cardiac tissue suggest that the production of oxygen free radicals is accelerated by an ischemic injury. Based on the study, we believe that the cardiac tissue is particularly susceptible to the effects of ischemia and reperfusion injury. Damage to cardiac tissue is probably the major cause of mortality following acute ischemic injury in a limb.

Animals↗

PubMed1999

Cytokine and prostaglandin E2 release from leukocytes in response to metal ions derived from different prosthetic materials: an in vitro study.

Cytokines produced by leukocytes in the periprosthetic membranes surrounding joint replacements have been implicated as causal agents in osteolysis and prosthetic loosening. In this study, we used an in vitro leukocyte culture system to monitor the response of leukocytes to various metal ions and their possible roles in the mechanism of aseptic loosening. Human peripheral leukocytes were isolated and incubated with various concentrations of Co2+, Cr3+, and Ti3+ ions. Leukocyte cell counts and the levels of the tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), interleukin-6 (IL-6) and prostaglandin E2 (PGE2) released into the media were analyzed at 1 h, 3 h, and 1, 3, and 7 day intervals. The results showed that adding different metal ions into leukocyte cultures did not affect the cell counts. Exposure of leukocytes to Co2+ ion increased the release of TNF-alpha, IL-6, and PGE2. Exposure of leukocytes to Cr3+ ion did not increase the release of TNF-alpha but increased the secretion of IL-6 and PGE2. In contrast, exposure of the leukocytes to Ti3+ ions was associated with a decrease in the release of TNF-alpha and PGE2 and a minimal change in IL-6 noted after 7 days' culture. The present study elucidated the possible mechanisms involved in periprosthetic osteolysis and the inflammatory response of human leukocytes to metal ions. We found that cobalt ion is the most potent stimulant for cytokines and prostaglandin secretion by leukocytes. This elucidation, in combination with other efforts to reduce the generation of wear debris and metal ions, may improve the longevity of orthopedic implants.

Cell Count↗

PubMed1999

Thermal decomposition and reconstruction of hydroxyapatite in air atmosphere.

In this paper, the decomposition and reconstruction behavior of hydroxyapatite (HAP) during heating and cooling in air atmosphere were studied. The commercial HAP were chosen and gradually heated to 1500 degrees C and cooled to room temperature by a program controlled SiC heated furnace. X-ray diffractometer (XRD) and Fourier-transformed infrared (FTIR) analysis were used to investigate the change of crystalline phases and functional groups of HAP at different temperatures. Weight change of samples was recorded by thermogravimetric analysis (TGA) during heating and cooling. The results revealed that HAP would gradually release its OH- ions and transform into OHAP in the temperature of 1000-1360 degrees C. Above 1360 degrees C, the OHAP would decompose into TTCP and alpha TCP phase. The OH- stretching bands of HAP could be traced by FTIR even at the temperature of 1350 degrees C which was the eve of HAP decomposition. It reflected that the HAP did not dehydrate completely before decomposed. We speculated that some of OAP might be formed during dehydration and there were a great amount of OHAP still left in the system even up to the temperature of decomposition. In the temperature of 1400-1500 degrees C, there were no significant different in XRD patterns, only TTCP and alpha TCP crystalline phase were observed. When the HAP gradually cooled down from 1500 degrees C, a part of TTCP and alpha TCP would directly reconstruct into oxyapatite (OAP) around 1350 degrees C. OAP existed in the temperature of 1350 to 1300 degrees C during cooling. When the temperature down to 1290 degrees C, a part of TTCP and alpha TCP reconstructed into OHAP by rehydration reaction and OAP were rehydrated into OHAP as well. At 1100 degrees C, the rest of TTCP and alpha TCP would reconstitute into HAP. As the temperature decreased, the OHAP were gradually rehydrated and reconstituted into HAP.

Biocompatible Materials↗

PubMed1998

Triple helix formation by (G,A)-containing oligonucleotides: asymmetric sequence effect.

Sequence effects on the stability of purine-motif (also called (G, A)-motif) triple helix have been investigated through two symmetry-related systems: one of them had a 5'(GGA)43' core sequence of triplex-forming oligonucleotides (TFOs), whereas the other one had a reversed 5'(AGG)43' core sequence. These (G,A)-containing TFOs were prone to self-associate into intermolecular complexes at room temperature. The competition of TFOs' self-association with triple helix formation was assessed, and minimized. By varying the lengths and the terminal base sequences of TFOs, the following were found that (1) The stability of two triple helices with identical length and base composition but reverse strand orientation may be significantly different (up to a factor of 6). (2) When the 5'(GGA)43' core sequence was extended at the 3'-end by a G, the 13-nt TFO exhibited 3- and 5-fold higher affinity toward the target double-stranded DNA (dsDNA) than the longer 14-nt and 15-nt TFOs in which one and two A(s) were added at the 3'-end of the 13-nt TFO, respectively. In contrast, when the similar extensions occurred at the 5'-end of the 5'(AGG)43' core sequence, the length increase provided a higher binding affinity of TFOs toward the target duplex. (3) The nature of the base triplets involved at the ends of triple helices may have great influence on triplex stability. The observed asymmetric sequence effect of the (G,A)-motif triple helix formation is discussed in terms of the binding strength of the first base triplet(s) at the 3' end which seems to be deeply involved in the nucleation step of triple helix formation and therefore to be a determining factor for triplex stability.

Adenine↗

PubMed1998

Optimization of alternate-strand triple helix formation at the 5'CpG3' and 5'GpC3' junction steps.

Oligonucleotide-directed triple helix formation normally requires a long tract of oligopyrimidine.oligopurine sequence. This limitation can be partially overcome by alternate-strand triple helix (or switch triple helix) formation which enables recognition of alternating oligopurine/oligopyrimidine sequences. The present work is devoted to the optimization of switch triple helix formation at the 5'CpG3' and 5'GpC3' junction steps by combination of base triplets in Hoogsteen and in reverse Hoogsteen configurations. Rational design by molecular mechanics was first carried out to study the geometrical constraints at different junction steps and to propose a "switch code" which would optimize the interactions at junctions. These predictions were further checked and validated experimentally by gel retardation and DNase I footprinting assays. It was shown that the choice of an appropriate linker nucleotide in the switching third strand plays an important role in the interaction between oligonucleotides and alternating oligopurine/oligopyrimidine target sequences at different junctions: (i) the addition of a cytosine at the junction level in the oligonucleotide optimizes the crossover at the 5'CpG3' junction, whereas (ii) the best crossover at the 5'GpC3' junction step is achieved without any additional nucleotide. These results provide a useful guideline to extend double-stranded DNA sequence recognition by switch triple helix formation.

Base Composition↗

PubMed1998

Triple helices formed at oligopyrimidine*oligopurine sequences with base pair inversions: effect of a triplex-specific ligand on stability and selectivity.

Oligonucleotide-directed triple helix formation is mostly restricted to oligopyrimidine*oligopurine sequences of double helical DNA. An interruption of one or two pyrimidines in the oligopurine target strand leads to a strong triplex destabilisation. We have investigated the effect of nucleotide analogues introduced in the third strand at the site opposite the base pair inversion(s). We show that a 3-nitropyrrole derivative (M) discriminates G*C from C*G, A*T and T*A in the presence of a triplex-specific ligand (a benzo[e]pyridoindole derivative, BePI). N6-methoxy-2,6-diaminopurine (K) binds to an A*T base pair better than a T*A, G*C or C*G base pair. Some discrimination is still observed in the presence of BePI and triplex stability is markedly increased. These findings should help in designing BePI-oligonucleotide conjugates to extend the range of DNA sequences available for triplex formation.

Chromosome Inversion↗