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Biomedical subjects

C P Lin

Publications and source records attributed to C P Lin.

At least 109 records · Page 6Linked to original sources

Picosecond neodymium:yttrium lithium fluoride laser sclerectomy.

The picosecond neodymium:yttrium lithium fluoride laser, a high-power, short-pulse laser that uses low energy per pulse and has a high repetition rate and a highly coherent energy source of 1,053 nm, was used to perform sclerectomies by an ab externo approach in human cadaver eyes. We evaluated laser settings with regard to full-thickness scleral perforation and thermal damage to surrounding tissue. We studied energy per pulse, depth per burst (a present number of spots), spot and line separation of the pattern (spacing between spots of the laser in length [spot separation] and width [line separation]), and total energy required to perforate the sclera. Efficiency was determined by evaluating which settings required the fewest spots and least total energy to perforate the sclera. We studied histologic sections of the sclerectomy sites to determine thermal damage to the surrounding sclera. The picosecond neodymium:yttrium lithium fluoride laser is effective in performing full-thickness sclerectomy with minimal thermal damage to the surrounding tissue. The anterior chamber could be penetrated with an average total energy of 13.3 +/- 0.4 (SEM) J. The sclerectomy size was 545 +/- 11 microns externally and 163 +/- 4 microns internally. Successful sclerectomies were performed with as little as 3 to 5 microns of thermal damage to the surrounding scleral tissue with 250 microJ per pulse.

Cadaver↗

Scanning electron microscopy of type I collagen at the dentin-enamel junction of human teeth.

The dentin-enamel junction constitutes a unique boundary between two highly mineralized tissues with very different matrix composition and physical properties. The nature of the boundary between the ectoderm-derived enamel and mesoderm-derived dentin is not known. This study was undertaken to identify the presence, type, and distribution of collagen at the dentin-enamel junction as an initial step in understanding its structural-functional role in dental occlusion. Sections of human teeth were demineralized with 0.1 M neutral EDTA and examined by high-resolution field-emission scanning electron microscopy at low accelerating voltage. Enamel and dentin were observed to be linked by many parallel 80-120-nm diameter fibrils, which were inserted directly into the enamel mineral and also merged with the interwoven fibrillar network of the dentin matrix. Immunogold labeling for collagen was visualized by secondary electron imaging and backscatter electron imaging at low accelerating voltage. The collagen fibrils at the junctional zone as well as in the dentin matrix were identified as Type I collagen. Collagenase digestion led to loss of the fibrillar structures and prevented immunogold labeling with antibody specific to Type I collagen. Consequently, the dentin-enamel junction can be regarded as a fibril-reinforced bond which is mineralized to a moderate degree.

Collagen↗

A tetrazolium-based(MTT) assay for measuring viability of cultured porcine corneal endothelium.

A tetrazolium-based(MTT) assay was used to evaluate the viability of cultured porcine monolayer corneal endothelial cells. In this study, we tried to find out the best spectra at which scanning multiwell ELISA spectrophometer can obtain the widest range of absorbance (optical density, O.D.) and the ranges of numbers of corneal endothelial cells in which absorbance is in a significant linear relationship with cell numbers. Among the three tested spectra of 450, 540, and 630nm, the widest range of absorbance was at 540 nm, from 0.1193 to 0.6183. At all three tested spectra, numbers of corneal endothelial cells from 50 to 30,000 were in a significant linear relationship with absorbance. Therefore, in the future studies to assay the viability of corneal endothelial cells, we shall choose the spectrum of 540 nm for reading with a scanning multiwell ELISA spectrophotometer, and absorbance(O.D.) from 0.1193 to 0.6183 will be accepted, representing cell numbers from 50 to 30,000.

Animals↗

Effect of dextran on predamaged corneal endothelium: an organ culture study.

Dextran T500 was used as a supplement in organ culture medium to evaluate its effect on corneal endothelium with disseminated damage. After 1 week of culturing, the control group showed an intact endothelium throughout the entire cornea. However, in the dextran-supplemented group, multiple endothelial necroses were observed in 5 of 10 corneas. Besides, intracytoplasmic vacuoles and granules, deposits on the endothelial surface and widened intercellular spaces were quite numerous. The potential toxicity of dextran to degenerated corneal endothelial cells may be responsible for these findings.

Animals↗

A simultaneous bilateral attack of infectious corneal ulcers in an extended-wear soft contact lens wearer: a case report.

A case study was made of a 23-year-old female with simultaneous bilateral infectious corneal ulcers related to extended-wear soft contact lenses. Cultures revealed Pseudomonas aeruginosa and Acinetobacter calcoaceticus. Multiple high risk factors were combined, including extended-wear, poor lens care, bilateral alternate wearing, and smoking. Amphetamine addiction may also have played a role. After treatment, a visual acuity of 20/20 was achieved in both eyes by the use of rigid gas-permeable contact lenses.

Adult↗

Detection of bcr/c-abl mRNA in chronic myelogenous leukemia by polymerase chain reaction to identify chromosome translocation.

The hallmark of chronic myelogenous leukemia (CML) is the Philadelphia chromosome (Ph1) which is caused by a translocation of the c-abl gene from chromosome 9 to the breakpoint cluster region (bcr) on chromosome 22. Polymerase chain reaction (PCR) can be used to detect the chimeric bcr/c-abl mRNA as evidence of the translocation. We applied a very simple and quick method of isolating cytoplasmic RNA, as well as reverse transcription and PCR in detecting bcr/c-abl mRNA of seven CML patients in various stages. Our results showed that only a small amount of either peripheral blood or bone marrow material was required for the expression of the bcr/c-abl mRNA. This is a very fast and time-saving method since a one-step method of cytoplasmic RNA isolation is used instead of the several steps in total RNA isolation as published in other literature.

Base Sequence↗

Detection of hemoglobin E heterozygotes by using polymerase chain reaction and direct DNA sequencing: report of a case.

From an overseas Chinese born in Thailand, an extraordinarily high level of A2 + E band (20.2%) was found during routine cellulose acetate Hb electrophoresis. With a suspicion of Hb E, the beta-globin gene was studied by using the polymerase chain reaction (PCR) and direct DNA sequencing method. A 1.4 kb fragment covering the whole beta-globin gene was amplified by PCR. Sequence analysis of the amplified product revealed a GAG > AAG transversion at codon 26, which resulted in an amino acid substitution of lysine for glutamic acid. A normal sequence at the corresponding codon was noted in the other allele; hence, this patient is a heterozygote of Hb E. PCR and direct DNA sequencing provide a rapid method for detection of Hb E from a small amount of DNA.

Adult↗

Rapid prenatal diagnosis of Hb Bart's hydrops fetalis in southeast Asia area by polymerase chain reaction.

We used the polymerase chain reaction (PCR) to amplify the breakpoint area of alpha-thalassemia-1 of Southeast Asia type and several parts of the alpha-globin gene cluster to make a differential diagnosis between alpha-thalassemia-1 and Hb Bart's hydrops fetalis. The procedure involved three primers to detect the homozygote of alpha-thalassemia-1, then amplifies the other alpha-globin gene cluster with three other pairs of primers to double check the results. The PCR products were checked again by allele specific probes. Twenty-two cases were diagnosed prenatally, two were normal, 17 were alpha-thalassemia-1, and three Hb Bart's hydrops fetalis. All cases were confirmed either by Southern blot hybridization or follow-up by sonography or after delivery. No false positive or false negative results were obtained by our strigent procedure. We conclude it to be a rapid, accurate and economic method.

Asia, Southeastern↗

Optical coherence tomography.

A technique called optical coherence tomography (OCT) has been developed for noninvasive cross-sectional imaging in biological systems. OCT uses low-coherence interferometry to produce a two-dimensional image of optical scattering from internal tissue microstructures in a way that is analogous to ultrasonic pulse-echo imaging. OCT has longitudinal and lateral spatial resolutions of a few micrometers and can detect reflected signals as small as approximately 10(-10) of the incident optical power. Tomographic imaging is demonstrated in vitro in the peripapillary area of the retina and in the coronary artery, two clinically relevant examples that are representative of transparent and turbid media, respectively.

Coronary Disease↗

Micron-resolution ranging of cornea anterior chamber by optical reflectometry.

Noncontact, high resolution measurements of anterior eye structures using optical coherence domain reflectometry are described. Distances between intraocular structures are measured by directing a beam of short coherence length light onto the eye and performing an interferometric measurement on the optical group delay of reflected signals. Measurements of corneal thickness, corneal excision depth, and anterior chamber depth are demonstrated in vitro, and the location of tissue boundaries is resolved to within +/- 2 microns. The full-width-half-maximum longitudinal resolution is 10 microns. Sensitivities to reflected signals as small as 10(-10) of the incident power are achieved by heterodyne detection.

Animals↗

The effects of epidermal growth factor and chondroitin sulfate on the animal corneal endothelial cell culture.

In order to investigate the effects of different culture media containing various concentrations of epidermal growth factor (EGF) and chondroitin sulfate (CDS) on the growth of cultured corneal endothelial cell, pig corneal endothelial cells were used for this study. The cells were divided into 7 groups and each group was cultured with a medium containing: 1. Eagle's minimal essential medium with Earle's salt (EMEM) only; 2. EMEM + EGF (10 ng/ml); 3. EMEM + EGF (100 ng/ml); 4. EMEM + CDS (1 mg/ml); 5. EMEM + CDS (25 mg/ml); 6. EMEM + EGF (10 ng/ml) + CDS (1 mg/ml); 7. EMEM + EGF (100 ng/ml) + CDS (25 mg/ml), respectively. The results are shown below: (1) High concentration of EGF (100 ng/ml) stimulated the growth of pig corneal endothelial cells, shortened doubling time and the cells reached confluence earliest in group 3. But a low concentration of EGF (10 ng/ml) showed no effects on the growth of pig corneal endothelial cells. (2) A high concentration of CDS (25 mg/ml) might retard the growth of pig corneal endothelial cell, but a low concentration of CDS (1 mg/ml) has no retarding effects on the growth of pig corneal endothelial cells. (3) A high concentration of EGF could "antagonize" the CDS induced growth retarding effects of endothelial cells. (4) Those cells cultured with a medium containing high concentrations of EGF had more mitotic activity, including many prominent binuclear and polynucleolar cells. (5) The cells cultured with a medium containing high concentrations of CDS showed a more flat-shaped morphology under observation with a phase contrast microscope.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[A self-made corneal graft marker made from disposable trephine].

A homemade corneal marker was contrived from a Pharmacia Superblade disposable corneal trephine by cutting away one arm of the trephine and then bending the single remaining arm to an angle of 45 degrees to the handle. The radii of the trephines ranged from 6.5mm to 8.5mm, at successive increments of 0.25mm. In optical corneal grafts, this marker can be used to pinpoint the center of the cornea, thus providing more accurate centralization of the area to be grafted. In therapeutic corneal grafts, the marker provides a more accurate determination of the most suitable area to be grafted. Unnecessary removal of healthy tissue or incomplete removal of diseased tissue are thus avoided. This marker is made easily and at no additional expense, and show itself to be a useful instrument to accurately determine the graft area in corneal surgery.

Corneal Transplantation↗

Effects of fibroblast growth factor and chondroitin sulfate on predamaged corneal endothelium. An organ culture study.

Fibroblast growth factor (FGF) and chondroitin sulfate (CDS) were used as supplements in organ culture medium to compare the regeneration ability of corneal endothelium with scattered damages. After 1 week of culturing, cell densities in both FGF-supplemented and FGF + CDS-supplemented groups were not higher than the densities of the control groups. Cells in both groups showed polymorphism. In both CDS-supplemented groups, the corneas were thinner. The cell density in the group supplemented with only CDS was higher than that of the control group, but in the CDS + FGF-supplemented group the cell density was not higher than that of the control and the morphology was even worse. Combining of FGF and CDS in culture medium appeared to be disadvantageous.

Animals↗

The role of serum-supplements in organ culture for pre-damaged corneal endothelium.

Thirty-six pig corneas were shaken in 0.18% NaCl to induce disseminated corneal endothelial damage. After treatment, the corneas were divided and cultured in 3 different combinations of media to study the effects of serum-supplements in corneal endothelial regeneration. There were MEM+10% FCS (FCS-group; n = 10); MEM+10% Nu-Serum (Nu-group; n = 10) and MEM only (control group; n = 16). After one week of culturing, most of the corneas in all 3 groups, despite the pre-damage, kept intact endothelia. This means that the endothelia were repaired whether the sera were supplemented or not. The cell densities of each were 2,454 +/- 450 cells/mm2, 2,776 +/- 476 cells/mm2 and 2,627 +/- 558 cells/mm2 respectively. The differences between each group were statistically insignificant. The morphological findings were also without much difference. When considering the risk of serum induced hypersensitivity, FCS can be replaced by Nu-Serum in the organ culture medium for corneal preservation. Since the advantage of serum-supplements in medium-term organ cultures seems limited, the necessity of serum supplements should be further reevaluated.

Animals↗

High-speed photography of Er: YAG laser ablation in fluid. Implication for laser vitreous surgery.

The mechanism of Er:YAG laser-induced long-range damage in intraocular surgery was investigated using high-speed photography. A short pulse of 2.94-microns radiation delivered by an optical fiber into an aqueous medium causes rapid localized heating and vaporization and creates a bubble at the tip of the fiber. The size of the bubble depends on the pulse energy and is about 1 mm at 1 mJ. The shape of the bubble has multiple lobes, which can be attributed to the spiky output of the laser pulse. The expanding bubble can cause thermal and mechanical damage to tissues. In addition, laser spikes propagating through the bubble can strike and damage tissue on the distal side of the bubble. In both mechanisms the damage zone approximates the bubble size and can be greater than 1 mm, ie, 1000 times the steady-state absorption length of water at 2.94 microns. The authors discuss ways to reduce the damage zone by bubble confinement.

Hot Temperature↗

Detection of beta-globin gene from single hairs.

DNA can be extracted from hair shafts and hair roots. The content of DNA in hairs is usually limited: the root end of hairs may contain 0.5 micrograms DNA and shed hairs contain less than 10 ng DNA. DNA analysis with restriction fragment length polymorphism (RFLP) requires microgram amounts of DNA. Such DNA cannot be obtained from such samples as single hairs or blood stains. Even with a little amount of DNA, specific genes of DNA can be greatly amplified to more than 10(6)-fold in vitro by polymerase chain reaction (PCR). We extracted the DNA from a half, 1, and 2 hair roots and 4 hair shafts and amplified the DNA with primer pairs (5'-GCACCATTCTAAAGAATAAC-3', 5'-GGATTGTAGCTGCTATTAGC-3') of the beta-globin gene, covering part sequences of the IVS-2 region by using the polymerase chain reaction for 50 cycles. The electrophoresis of the PCR product revealed a 131 base pairs band. Finally, we hybridized the PCR product with an IVS-2 probe (5'-GGGTTAAGGCAATAGCAAT-3'). A fragment of the IVS-2 extending from nucleotide 612 to 742 of the beta-globin gene can be demonstrated in the slot blot filter of hair roots and hair shafts, but not in the 1 microgram of blood which was not amplified by PCR. Detection of the beta-globin gene from single hairs by using the PCR may be useful for the diagnosis of thalassemia from single hairs.

Base Sequence↗