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Biomedical subjects

Y C Tsai

Publications and source records attributed to Y C Tsai.

At least 19 recordsLinked to original sources

Contiguous patches of normal human mammary epithelium derived from a single stem cell: implications for breast carcinogenesis.

Tissue clonality can be assessed in females by analyzing the methylation status of polymorphic DNA markers on X-linked genes because extensive de novo methylation of one allele at the preimplantation stage is associated with its permanent inactivation. We applied X chromosome inactivation toward understanding human breast morphogenesis by examining the nonmalignant breast epithelium from two reduction mammaplasties and a mastectomy. We found that entire lobules and large ducts of normal breast tissue have the same X chromosome inactivated, suggesting that they are derived from the same stem cell. The regions of inactivation of a particular X chromosome do not extend over an entire breast, so that ducts and lobules with opposite chromosomes inactivated are present within a single breast. Potential relevance of these observations for malignant transformation is discussed.

Base Sequence

Intratumoral blood flow in uterine myoma correlated with a lower tumor size and volume, but not correlated with cell proliferation or angiogenesis.

OBJECTIVE: To investigate the correlation of intratumoral blood flow in uterine myoma with cell proliferation, angiogenesis, tumor size, and tumor volume. METHODS: Thirty-nine patients who had been scheduled for surgery because of symptomatic uterine myomas were evaluated by transvaginal sonography and color Doppler ultrasound before surgery. The largest dimension of each tumor and the volumes of myomas were determined ultrasonographically. Pulsatility index (PI) was determined by color Doppler ultrasound according to the maximum systolic, end-diastolic, and the mean flow velocities measured within the uterine nodules. After surgery, the paraffin-embedded slides containing representative leiomyoma tissues were stained with hematoxylin and eosin, proliferating cell nuclear antigen for measurement of cell proliferation, and factor VIII for quantitation of microvessel density. The ultrasonographic findings were correlated postoperatively with pathologic findings, and the data were analyzed by simple linear regression and Fisher r to z transformation. RESULTS: Simple regression analysis of the intratumoral PI values on the sizes of myomas showed a negative correlation (r = -0.47, P = .003; n = 39), whereas a less significant correlation between PI values and tumor volumes was observed (r = -0.42, P = .008). In contrast, no statistically significant correlation was observed between the intratumoral PI values and the values of the proliferating cell nuclear antigen index (r = 0.10, P = .547) or microvessel density counts (r = 0.18, P = .282). CONCLUSION: The intratumoral blood flow by transvaginal color Doppler ultrasound correlated with a reduced tumor size and tumor volume, but did not correlate with cell proliferation or angiogenesis.

Adult

A comparison of the effect of halothane on N-methyl-D-aspartate and non-N-methyl-D-aspartate receptor-mediated excitatory synaptic transmission in the hippocampus.

Halothane depresses synaptic transmission in the rat brain. First we determined the concentration of halothane which decreased the amplitude of the population spike recorded in the CA1 region of the hippocampus to 50% of the control value (105 +/- 4.9 micrograms/mL [0.53 mM] halothane). Hippocampal glutamate receptors are divided into N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole proprionate (AMPA) and kainate (non-NMDA) subtypes. The NMDA and non-NMDA receptors were blocked with (+/-)-2-amino-5-phosphonopentanoic acid (AP5) (30 microM), and 6,7-dinitroquinoxaline-2,3-dione (DNQX) (10 microM), respectively, to allow observation of the effects of halothane on the NMDA and non-NMDA receptors, respectively. gamma-Aminobutyric acid type A (GABAA) receptors were blocked in all studies with picrotoxin (PTX) (40 microM). When the non-NMDA receptors were blocked a halothane concentration of 38.1 +/- 5.6 mg/mL was required to produce a further 50% decrease in population spike amplitude. When NMDA receptors were blocked with AP5 or only GABAA receptors were blocked the halothane concentrations needed to produce 50% block were higher than needed for the control (160.8 +/- 17.8 and 190.2 +/- 12.1 microgram/mL, respectively). These studies indicate that the NMDA receptors are more sensitive to the effects of halothane than the non-NMDA receptors.

2-Amino-5-phosphonovalerate

Mosaicism in human epithelium: macroscopic monoclonal patches cover the urothelium.

Previous studies of chimeric animals and human tissues have shown the clonal nature of organ development, giving clues as to the normal development of organs and also to abnormal developments, such as atheromatous plaques. The clonal nature of bladder cancer in female patients has been demonstrated, but little has been known of the clonal development of the normal urothelium. Using an X chromosome inactivation analysis of cells microdissected from histologic slides from the female human bladder, macroscopic urothelial patches of monoclonality were detected. These patches are about 120 mm.2 in size, contain about 2 x 10(6) cells each and reflect the presence of coherent cellular families composed of stem cells and their differentiated derivatives. The large size of these patches was surprising when compared with previously reported patch sizes in other organ systems. The patches most probably are composed of the descendants of the original founder cells, which would suggest that only 200 to 300 cells participated in the formation of the urothelium. The limited number of stem cells, each giving rise to millions of cells may provide an explanation for the "field defect" that is often referred to in the pathogenesis of bladder cancer, as different cell patches may possess different predispositions to tumorigenesis.

Dosage Compensation, Genetic

Production and characterization of keratinase of a feather-degrading Bacillus licheniformis PWD-1.

The keratinase produced by Bacillus licheniformis PWD-1 was induced by feather powder. Maximal enzyme production could be achieved by culturing in a medium containing 1% hammer-milled feather powder (100 mesh) at 45 degrees C for 30 h. Maximal growth of PWD-1 was achieved at 50 degrees C, and maximal enzyme induction was at 45 degrees C. The molecular mass and isoelectric point of this enzyme were 31.4 kDa and 8.5, respectively. This enzyme was stable from pH 5 to 12. The optimal reaction pHs for feather powder and casein were 8.5 and 10.5 to 11.5, respectively. The optimal reaction temperature was 50 degrees C to 55 degrees C. The relative activity of this enzyme toward casein, feather powder, keratin, elastin, and collagen was 100:52:41:18:7, and 100:56:32:3 for Suc-AAPL-pNA, Suc-AAPF-pNA, Suc-AAPM-pNA, and Suc-AAVA-pNA (Suc, succinyl; pNA, p-nitrophenylanilide).

Amino Acid Sequence

Cerebral microvascular reactivity to carbon dioxide during isoflurane anesthesia as assessed by laser-Doppler flowmetry.

BACKGROUND: To determine the effects of carbon dioxide on the cerebral microcirculation during isoflurane anesthesia. METHODS: We studied 10 rabbits using laser Doppler flowmetry on the parietal cortex. After surgical preparation, the end-tidal concentration of isoflurane was maintained at one MAC (2.05%). Mean arterial pressure, rectal temperature and hematocrit were kept constant. The arterial carbon dioxide tension (PaCO2) was randomly adjusted to three levels: 20-25 mmHg (hypocapnia); 35-40 mmHg (normocapnia); and 45-50 mmHg (hypercapnia). Cerebral blood flow (CBF) was measured after target PaCO2 had been achieved. RESULTS: CBF at normocapnia was 27.4 +/- 13.5 (mean +/- SD) ml/100 gm/min, which increased to 42.4 +/- 14.3 ml/100 g/min (p < 0.01) and decreased to 15.3 +/- 7.5 ml/100 g/min (p < 0.05) at hypercapnia and hypocapnia respectively. The slopes of CBF versus PaCO2 were 1.60 +/- 0.63 ml/100 g/min/mmHg from normocapnia to hypercapnia and -0.79 +/- 0.51 ml/100 g/min/mmHg from normocapnia to hypocapnia. The corresponding percentage of alteration in CBF on the respective change of PaCO2 was 7.2 +/- 5.1 and -3.0 +/- 1.2%/mmHg. CONCLUSIONS: These data indicate that the changes of the cerebral microvascular blood flow reflect the CO2 effects during isoflurane anesthesia.

Anesthesia

Predictive value of endometrial sonography in ovulation induction for intrauterine insemination.

This prospective study was undertaken to evaluate the prognostic value of the endometrial sonographic pattern on the day of human chorionic gonadotropin (hCG) administration in patients who underwent intrauterine insemination. The endometrial sonographic patterns were classified as homogenous, hyperechogenic (A), intermediate isoechogenic (B) and multiple-layered "triple-line" (C). Type A had no pregnancies, whereas type B had a 20% fecundity rate and type C had a 21.1% fecundity rate. The continuing pregnancy rate was 10% in type B, as compared to 19.3% in type C. The data suggest that the endometrial sonographic pattern on the day of hCG administration may be of predictive value regarding fecundity and continuing pregnancy in the intrauterine insemination cycle.

Adult

Cardiotoxin II from Taiwan cobra venom, Naja naja atra. Structure in solution and comparison among homologous cardiotoxins.

The three-dimensional structure in solution of cardiotoxin II, a membrane toxin from the venom of Taiwan cobra, Naja naja atra, was determined using 1H nuclear magnetic resonance spectroscopy and molecular modeling based on the hybrid distance geometry/dynamic simulated annealing technique. A complete sequence-specific proton assignment was obtained, and the secondary structures of the protein were determined from information on nuclear Overhauser effect connectivities, coupling constants, and hydrogen exchange were confirmed using the main-chain-directed strategy. Twelve simulated annealing structures found to be within a single family were selected based on the condition of distance constraint violation less than 0.02 nm and the dihedral angle violation less than 4 degrees. The average atomic root mean square deviation between the selected structures and their geometric average are 0.079 nm for the backbone atoms and 0.137 nm for all heavy atoms; they are 0.044 nm and 0.117 nm, respectively, when considering the secondary structural residues only. The molecule adopts a compact structure consisting of three major loops emerging from a globular head. These loops contain five strands to form double- and a triple-stranded antiparallel beta sheets. Comparisons are made between this structure and those of its homologous cardiotoxins in order to derive further information on their structural variations.

Amino Acid Sequence

Hyponatremia and syndrome of inappropriate antidiuretic hormone secretion in adult spinal surgery.

STUDY DESIGN: Patients undergoing spinal surgery were monitored for sodium balance, fluid type, and volume input and output during surgery and for the first 3 postoperative days. OBJECTIVE: To prospectively document the true incidence of the syndrome of inappropriate antidiuretic hormone secretion and hyponatremia, and identify risk and protective factors for the development of the syndrome of inappropriate antidiuretic hormone secretion after spinal surgery. METHODS: Data on medical history, surgical procedure, estimated blood loss, and volumes and types of intraoperative and postoperative fluids were collected on 116 consecutive spinal surgery patients during March to July 1992. RESULTS: One hundred one spinal operations in 96 patients were evaluated. There were 48 males and 48 females, with a mean age of 52 years (range, 16 to 90 years). Hyponatremia developed in 45 (44.6%) patients. The etiology of hyponatremia was the syndrome of inappropriate antidiuretic hormone secretion in seven patients (6.9%), hypovolemia in 19 patients (18%), and other causes in six patients. CONCLUSIONS: Spine patients are at risk for hyponatremia and the syndrome of inappropriate antidiuretic hormone secretion. The incidence of the syndrome of inappropriate antidiuretic hormone secretion was 6.9%. Serum sodium should be monitored postoperatively. Patients who undergo a revision operation have an approximately two to four times greater risk of being affected by the syndrome of inappropriate antidiuretic hormone secretion than those who have primary surgery.

Blood Loss, Surgical

Two molecular pathways to transitional cell carcinoma of the bladder.

Noninvasive transitional cell carcinomas of the bladder can have two distinct morphologies suggesting they contain different genetic alterations. Papillary transitional cell carcinomas (T(a) tumors) are often multifocal and only occasionally progress, whereas flat tumors (carcinomas in situ, CIS), frequently progress to invasive disease. We examined 216 bladder tumors of various stages and histopathologies for two genetic alterations previously described to be of importance in bladder tumorigenesis. Loss of heterozygosity of chromosome 9 was observed in 24 of 70 (34%) T(a) tumors but was present in only 3 of 24 (12%) CIS and dysplasia lesions (P = 0.04). In contrast, only 1 of 36 (3%) T(a) tumors contained a p53 gene mutation compared to 15 of 23 (65%) CIS and dysplasias (P < 0.001), a frequency comparable to that observed in muscle invasive tumors (25 of 49; 51%). The presence of p53 mutations in CIS and dysplasia could explain their propensities to progress since these mutations are known to destabilize the genome. Analysis of several tumor pairs involving a CIS and an invasive cancer provided evidence that the chromosome 9 alteration may in some cases be involved in the progression of CIS to more invasive tumors, in addition to its role in the initiation of T(a) tumors. However, the CIS and secondary tumor were found to contain different genetic alterations in some patients suggesting divergent progression pathways. Bladder carcinogenesis may therefore proceed through two distinct genetic alteration pathways responsible for generating superficial tumors with differing morphologies and pathologies.

Alleles

Cerebral effects of nitrous oxide during isoflurane-induced hypotension in the pig.

We have studied the effects of nitrous oxide on cerebral blood flow (CBF), cerebral blood flow velocity (CBFV) and intracranial pressure (ICP) during isoflurane-induced hypotension in 10 pigs. CBF was measured using laser Doppler flowmetry, CBFV in the right middle cerebral artery was calculated using Doppler ultrasound and ICP was measured using an extradural ICP monitor. Each animal was studied under four conditions, examined sequentially: (i) mean intra-arterial pressure (MAP) 85 mm Hg, maintained with isoflurane, (ii) MAP 50-55 mm Hg, induced by isoflurane only, (iii) MAP 85 mm Hg, maintained with isoflurane and 50% nitrous oxide, and (iv) MAP 50-55 mm Hg, induced by isoflurane and 50% nitrous oxide. No significant differences were noted between conditions with respect to ICP. There was a significant difference in CBF during condition (ii) compared with (i) (mean 75(SD 21) vs 100(0)%) and during condition (iv) compared with (iii) (90(26) vs 109(13)%). Animals under condition (iv) exhibited a 20% reduction in CBFV compared with those under condition (iii) (57 vs 69 cm s-1). For animals under normotensive conditions, addition of nitrous oxide to isoflurane resulted in a 16% increase in CBFV (69 vs 60 cm s-1). Comparing isoflurane-induced hypotension ((ii) vs (iv)), there was no statistical difference in either CBF or CBFV on addition of 50% nitrous oxide. The correlation between changes in CBF and CBFV was not significant. We conclude that the use of nitrous oxide during isoflurane-induced hypotension has no significant effect on CBF, CBFV or ICP compared with the use of isoflurane alone.

Animals

Mutational spectrum in the p53 gene in bladder tumors from the endemic area of black foot disease in Taiwan.

An elevated risk of bladder cancer has been reported in the endemic region of 'black foot disease' on the southwest coast of Taiwan and may be related to high arsenic levels in artesian well water. Thirteen urothelial tumors from this endemic region were examined for mutations in exons 5-8 of the p53 gene to identify the effects of possible exogenous factors at the DNA level. DNA was extracted from archival tissue after microdissection of tumors and analyzed by PCR-SSCP (polymerase chain reaction-based single strand conformation polymorphism), followed by direct sequencing. Eight cases (62%) showed mutations and 9 of the 10 point mutations observed were transitions. The type and position of the mutations were not significantly different when compared with the spectra of p53 mutations previously reported for transitional cell carcinomas (TCCs). However, two of the mutations were CGC-->CAC base changes at codon 175, a mutational hotspot for many tumor types but previously unreported in TCCs except in cases associated with inflammatory agents. Three of the tumors examined were found to contain double mutations, a relatively rare mutagenic event in human cancers. Our results suggest that the agents responsible for the high risk of bladder cancer in the black foot disease region may operate through an inflammation-based mechanism which increases the amount of DNA damage per mutagenic event.

Adult

Purification and characterization of L-aminoacylase from Alcaligenes denitrificans DA181.

The L-aminoacylase produced intracellularly by Alcaligenes denitrificans DA181 was purified to homogeneity. This enzyme had an apparent molecular weight of 80,000, and was composed of two subunits of identical molecular weight. Its isoelectric point was pH 5.1. The optimal reaction temperature and pH were 65 degrees C and 8.0, respectively. This enzyme showed specificity toward N-acetyl-derivative of hydrophobic L-amino acids with N-acetyl-L-valine as the favored substrate, followed by N-acetyl-L-alanine.

Alcaligenes

Solution conformation of enopeptin A, a depsipeptide antibiotic, using 2D NMR and restrained molecular dynamics studies.

Studies on the solution conformation of the cyclic depsipeptide antibiotic enopeptin A have been carried out using 2D NMR and molecular modelling techniques. The proton resonances of the antibiotic in DMSO-d6 have been assigned by the use of TOCSY and ROESY experiments. The interproton distance information obtained from the ROESY experiments have been used as the basis for elucidating the probable structures in solution. The restrained molecular dynamics technique was applied to calculate the structures in solution, and six resultant structures with fewer distance constraint violations were obtained that satisfy the experimental restraints very well. The conformation of the cyclic moiety of the molecules is well defined whereas the aliphatic chain segment is disordered.

Amino Acid Sequence

Idiopathic autoimmune hemolytic anemia during pregnancy.

A case of idiopathic autoimmune hemolytic anemia occurring during the third trimester of pregnancy is presented. The hemolytic process was quite severe in the mother but responded to steroid therapy. A healthy baby without evidence of hemolysis was delivered uneventfully through the vagina. We postulate the mechanism of the discuss the clinical picture, management of the disease and the possible effect on the fetus.

Adult

The influence of focal ischemic brain injury on tail-flick latency in the rat.

Tail-flick latency (TFL), a measure of response threshold to pain, was investigated in rats with focal ischemic brain injury resulting from middle cerebral artery occlusion (MCAO) as compared to sham and non-operative (non-op) groups on the 1st, 7th, 14th, 21st and 28th days following surgery or anesthesia. No significant differences were observed between MCAO and sham groups postoperatively, indicating focal ischemic insult induced by this model had no significant influence on TFL. Increased sensitivity to TFL in early postoperative period was shown when compared sham or MCAO with non-op group. The correlation between TFL and the time from surgery was statistically significant on the 1st and 7th days in sham group and the 1st, 7th and 14th days in MCAO group. We conclude that the focal ischemic injury resulting from middle cerebral artery occlusion that has no obvious neurologic effects does not alter pain threshold; however, surgical procedure increases postoperative sensitivity to pain stimulation.

Animals

Propofol does not reduce infarct volume in rats undergoing permanent middle cerebral artery occlusion.

As propofol has been reported to have effects similar to those of thiopentone on cerebral blood flow, intracranial pressure and cerebral metabolic rate, we postulated that it might also have a protective effect on focal cerebral ischemia. This experiment was designed to test whether or not propofol can alter infarct volume resulting from permanent focal cerebral ischemia in rats. Forty-five rats were randomly assigned to three groups, all of which received xylaxine (5 mg kg-1 im) and ketamine (100 mg kg-1 ip) anesthesia. Rectal temperatures were controlled at 36-37 degrees C. A right temporal craniotomy was performed and the middle cerebral artery (MCA) was ligated with a 10-O suture. Both common carotid arteries (CCAs) were then occluded for 60 minutes using aneurysm clips. In Group 1 rats (n = 15), normal saline (1 ml kg-1 i.v.) was given right after MCA and CCAs ligation. In Group 2 rats (N = 15), propofol (10 mg kg-1 i.v. bolus) was given right after MCA and CCAs ligation. In Group 3 rats (n = 15), continuous intravenous infusion of propofol (16 mg kg-1 hr-1) was given for four hours following MCA and CCAs ligation, during which time mean arterial pressure was maintained approximately 90 mmHg. Twenty-four hours after ischemia, the brains were removed, frozen, sectioned and stained with 2% 2,3,5-triphenyltetrazolium chloride. The cross-sectional area of infarction on each brain slice was measured using computer imaging analysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals