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Biomedical subjects

K C Chen

Publications and source records attributed to K C Chen.

At least 19 recordsLinked to original sources

Computer evaluation of network dynamics models with application to cell cycle control in budding yeast.

Cellular processes are governed by complex networks of interacting genes and proteins. Theoretical molecular biologists attempt to describe these processes via mathematical models by writing biochemical reaction equations. Modellers are building increasingly larger and complex mathematical models to describe these cellular processes, making model evaluation a time consuming and difficult task. The authors describe an automatable process for model evaluation and a software system that implements this process. The software is adaptable to many types of models and is freely available along with all needed data files. The cell cycle control system for budding yeast is known in fine detail and constrained by more than 100 phenotypic observations in mutant strains. As an example, the authors apply their process to a model of cell cycle control in budding yeast containing dozens of regulatory equations and explaining nearly all of the known mutant phenotypes.

Cell Cycle↗

Cloning and expression of the genes associated with lipid metabolism in Tsaiya ducks.

Sterol regulatory element binding protein 1 (SREBP1) drives the expression of several lipogenic genes, whereas SREBP2 dictates the expression of every gene involved in cholesterolgenesis in mammals. In the current study, we cloned the cDNA fragments for SREBP1, SREBP2, fatty acid synthase (FAS), 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase), and very low density apolipoprotein-II (apoVLDL-II), the genes associated with lipid metabolism. Fifteen ducks immediately before the first egg was laid (18 wk old) and 15 ducks from the same population at an egg production rate of 80% were killed. Total RNA was extracted from liver and used to amplify the targeted genes by reverse transcription-PCR and screening of a cDNA library. The sequence data showed that Tsaiya duck SREBP1, SREBP2, FAS, and HMG-CoA reductase were highly homologous to that of chicken. Tsaiya duck SREBP1 mRNA was expressed in adipose tissue, cardiac muscle, skeletal muscle, liver, and ovary. The SREBP2 mRNA concentration was highest in liver and ovary. Concentrations of FAS and HMG-CoA reductase mRNA were high in liver and lower in other tissues. The apoVLDL-II mRNA was specifically expressed in the liver. The differences between mRNA concentrations of SREBP1, SREBP2, and FAS in the livers of laying and prelay ducks were not significant. However, the concentrations of hepatic HMG-CoA reductase and apoVLDL-II mRNA were higher in the laying ducks than in prelay ducks. Therefore, laying may affect particular aspects of lipid metabolism, especially biochemical pathways that involved apoVLDL-II and HMG-CoA reductase.

Adipose Tissue↗

Neurotrophic effects of vascular endothelial growth factor and neurotrophins on cultured major pelvic ganglia.

OBJECTIVE: To investigate the feasibility of using a ganglial culture system to screen various growth factors as potential therapeutic agents for pelvic nerve injuries. MATERIALS AND METHODS: The major pelvic ganglia (MPG) were isolated from male rats and attached to culture dishes with the aid of Matrigel (Becton Dickinson, Mountain View, CA, USA). Alternatively, the dorso-caudal region (DCR) of MPG, from which the cavernous nerves originate, was dissected and then attached to a Matrigel-coated coverslip. The MPG or DCR was cultured in serum-free medium supplemented with phosphate-buffered saline (PBS, control), 50 ng/mL of vascular endothelial growth factor (VEGF), 20 ng/mL of a neurotrophin (BDNF, NT3, or NT4), or combinations of these growth factors. After 2 days of incubation, the ganglial tissues with their outgrowing nerve fibres were stained for the expression of NADPH-diaphorase, tyrosine hydroxylase (TH) and acetylcholinesterase (AChE). The length and staining intensity of nerve fibres were analysed. RESULTS: The outgrowing fibres were significantly longer in MPG treated with any of the four tested growth factors than in PBS-treated MPG. The combination of VEGF and NT3 induced the best fibre growth. Improvements to the culturing conditions allowed a histological examination of the outgrowing fibres for the expression of nitric oxide synthase (NOS), TH and AChE. VEGF and BDNF were equally capable of inducing NOS- and TH-expressing fibres. BDNF was much weaker than VEGF for inducing AChE-expressing fibres. CONCLUSIONS: This improved culturing system is potentially useful for screening nerve-regenerating factors; VEGF had neurotrophic effects comparable with BDNF, NT3, or NT4.

Animals↗

Gene expression profiling of an arteriogenic impotence model.

Penile arterial insufficiency is one of the most common causes of ED. We have established a traumatic arteriogenic insufficiency rat model by the ligation of the pudendal arteries. To simulate both acute and chronic traumatic injuries, five ligation periods (6 h, 3 days, 7 days, 3 weeks, and 6 weeks) were chosen. By electrostimulation of the cavernous nerve, the intracavernous pressure was determined to be between 20 and 40-cm H(2)O for the ligated rats compared to around 100-cm H(2)O for the control rats. The erectile tissue in the corpus cavernosum of these rats was then subjected to microarray analysis, in which an array that contains cDNA fragments representing 1176 rat genes was used. The results demonstrated that normal rat corpus cavernosum expressed approximately 200 genes at detectable levels and that ligation produced differential expression of approximately 25 genes, depending on the duration of ligation. The most highly ligation-induced gene was apolipoprotein D (ApoD), with peak expression in the 3- and 7-day ligated rats. Three of the insulin-like growth factor binding proteins (IGFBP-1, 3, and 5) were upregulated in all ligated rats. IGFBP-6, which was one of the most highly expressed genes in the normal corpus cavernosum, was down-regulated in all ligated rats. Cysteine proteases of the cathepsin family were also differentially expressed between control and ligated rats, with cathepsin K being down-regulated most. A few genes were upregulated only in the 6-week ligated rats, including angiotensin-converting enzyme. Finally, VEGF, whose induction has been identified in many other ischemic tissues, was not induced in corpus cavernous tissue of ligated rats.

Animals↗

Vitamin D hormone confers neuroprotection in parallel with downregulation of L-type calcium channel expression in hippocampal neurons.

Although vitamin D hormone (VDH; 1,25-dihydroxyvitamin D(3)), the active metabolite of vitamin D, is the major Ca(2+)-regulatory steroid hormone in the periphery, it is not known whether it also modulates Ca(2+) homeostasis in brain neurons. Recently, chronic treatment with VDH was reported to protect brain neurons in both aging and animal models of stroke. However, it is unclear whether those actions were attributable to direct effects on brain cells or indirect effects mediated via peripheral pathways. VDH modulates L-type voltage-sensitive Ca(2+) channels (L-VSCCs) in peripheral tissues, and an increase in L-VSCCs appears linked to both brain aging and neuronal vulnerability. Therefore, we tested the hypothesis that VDH has direct neuroprotective actions and, in parallel, targets L-VSCCs in hippocampal neurons. Primary rat hippocampal cultures, treated for several days with VDH, exhibited a U-shaped concentration-response curve for neuroprotection against excitotoxic insults: lower concentrations of VDH (1-100 nm) were protective, but higher, nonphysiological concentrations (500-1000 nm) were not. Parallel studies using patch-clamp techniques found a similar U-shaped curve in which L-VSCC current was reduced at lower VDH concentrations and increased at higher (500 nm) concentrations. Real-time PCR studies demonstrated that VDH monotonically downregulated mRNA expression for the alpha(1C) and alpha(1D) pore-forming subunits of L-VSCCs. However, 500 nm VDH also nonspecifically reduced a range of other mRNA species. Thus, these studies provide the first evidence of (1) direct neuroprotective actions of VDH at relatively low concentrations, and (2) selective downregulation of L-VSCC expression in brain neurons at the same, lower concentrations.

Animals↗

Juvenile papillomatosis of the breast in a 9-year-old girl.

Juvenile papillomatosis (JP) is a rare benign, proliferative breast tumor in children. We observed a 9-year-old girl with a large soft-tissue mass at the lower outer quadrant of the left breast. Ultrasonography showed an ill-defined, inhomogeneous mass with numerous small, hypoechoic areas. The tumor was completely excised. Histopathology revealed JP of the breast. The case is presented and the literature is reviewed.

Breast↗

Purification and characterization of isoforms of beta-galactosidases in mung bean seedlings.

Five isoforms of beta-galactosidase (EC 3.2.1.23), designated as beta-galactosidases I-V, were isolated from five-day-old mung bean (Vigna radiata) seedlings. Beta-galactosidases II and III were purified to electrophoretic homogeneity by a procedure involving acid precipitation, ammonium sulfate fractionation, chromatography on diethylaminoethyl-cellulose (DEAE-Cellulose) and con A-Sepharose. and chromatofocusing. Beta-galactosidases I, II and III have the same molecular mass of 87 kDa. comprising two nonidentical subunits with molecular masses of 38 and 48 kDa, while beta-galactosidases IV and V have molecular masses of 45 and 73 kDa, respectively. All the enzymes were active against p-nitrophenyl-beta-D-galactoside, and to a lesser extent, p-nitrophenyl-alpha-L-arabinoside and p-nitrophenyl-beta-D-fucoside. The enzymes were inhibited by D-galactono-1,4-lactone, D-galactose, Hg2+, Ag+ and sodium dodecyl sulfate (SDS). Beta-galactosidases I, II and III were shown to be competitively inhibited by either D-galactono-1, 4-lactone or D-galactose. Isoforms I, II and III have a common optimal pH of 3.6, while isoforms IV and V have pH optima at 3.8 and 4.0, respectively. Isoelectric points of isoforms I, II and III were 7.7, 7.5 and 7.3, respectively. Double immunodiffusion analysis indicated that beta-galactosidases I, II, III and V are immunologically similar to each other, while beta-galactosidase IV shares partially identical antigenic determinants with the other four isoforms. The purified beta-galactosidases II and III were capable of releasing D-galactose residue from the hemicellulose fraction isolated from mung bean seeds.

Chromatography, Affinity↗

Epilepsy-induced decrease of L-type Ca2+ channel activity and coordinate regulation of subunit mRNA in single neurons of rat hippocampal 'zipper' slices.

L-type voltage-sensitive Ca2+ channels (VSCCs) preferentially modulate several neuronal processes that are thought to be important in epileptogenesis, including the slow afterhyperpolarization (AHP), LTP, and trophic factor gene expression. However, little is yet known about the roles of L-type VSCCs in the epileptogenic process. Here, we used cell-attached patch recording techniques and single cell mRNA analyses to study L-type VSCCs in CA1 neurons from partially dissociated (zipper) hippocampal slices from entorhinally-kindled rats. L-type Ca2+-channel activity was reduced by >50% at 1.5-3 months after kindling. Following recording, the same single neurons were extracted and collected for mRNA analysis using a recently developed method that does not amputate major dendritic processes. Therefore, neurons contained essentially full complements of mRNA. For each collected neuron, mRNA contents for the L-type pore-forming alpha1D/Ca(v)1.3-subunit and for calmodulin were then analyzed by semiquantitative kinetic RT-PCR. L-type alpha1D-subunit mRNA was correlated with L-type Ca2+-channel activity across single cells, whereas calmodulin mRNA was not. Thus, these results appear to provide the first direct evidence at the single channel and gene expression levels that chronic expression of an identified Ca2+-channel type is modulated by epileptiform activity. Moreover, the present data suggest the hypothesis that down regulation of alpha1D-gene expression by kindling may contribute to the long-term maintenance of epileptiform activity, possibly through reduced Ca2+-dependent AHP and/or altered expression of other relevant genes.

Animals↗

Duodenojejunal intussusception secondary to hamartomatous polyps of duodenum surrounding the ampulla of Vater.

Duodenojejunal intussusception is a rare pediatric emergency. A case of duodenojejunal intussusception secondary to hamartomatous polyps of the second portion of duodenum in a 10-month-old boy is reported. Surgical excision of the polyps and reduction of the intussusception were performed. Pathologic examination found hamartomatous polyps. This is the third case report of children in literature, but this is the first case of a child with intussusception surrounding the ampulla of Vater and a successful excision performed without damaging the ampulla of Vater.

Ampulla of Vater↗

Dynamics of changes in micronucleus frequencies in subjects post cessation of chronic low-dose radiation exposure.

To assess DNA damage remaining in peripheral lymphocytes, 48 individuals were evaluated twice for lymphocyte micronucleus frequencies by the cytokinesis-blocking cytochalasin B (CBMN) analysis post relocation from radio-contaminated apartments after various periods of time. The frequencies of CBMN at the first evaluation were significantly higher than those at the second examination (Chang et al., 1999c). These individuals were categorized into three groups: those with cumulative exposure of >300 mSv (defined as high exposure, HDose), those with 100-300 mSv (MDose) and those with <100 mSv (LDose). Using the Poisson mixed-effect model (Little et al., 1996), the estimated mean CBMN frequencies ( per thousand) for individuals in HDose, MDose and LDose exposure categories when they had only recently relocated were 21.8, 17.6 and 15.4, respectively. The estimated mean duration post relocation for the CBMN frequencies of these individuals to reduce to 10.2, the second CBMN frequency, on average, was 47.5, 37.2 and 28.3 months in the three exposure groups, respectively. The rates of change in CBMN frequencies were shown to be significantly higher in the HDose group than in the MDose and LDose groups. The results suggested a characteristic dose-dependent decline in the CBMN frequencies in the exposed population post cessation of chronic low-dose ionizing radiation exposure.

Adolescent↗

Legal and ethical considerations of assisted reproductive technology and surrogate motherhood in AOFOG countries.

With increasing use of assisted reproductive technologies (ART) and its related issues, the Ethics Committee of the Asia-Oceania Federation of Obstetrics and Gynecology (AOFOG), after discussion at its council meeting, took up the mission to understand the current legal and ethical aspects of this special subject in Asia-Oceania area before making proper recommendations. This Committee drafted a questionnaire to all 22 country member societies in 1995, and again conducted the same survey in 2000 to track changes in each country/region's laws and regulations. The results show that the current attitude and status of country member societies vary in a wide range and do not have formal legal codes in majority regulating the ethical aspects of ART. The use of ART and surrogacy is obviously affected by local traditions, popularity in the use of such techniques and cultural/religious concerns. The changes in 5 years are not major. Three countries do not as yet practice such techniques. However, liberalization of surrogate motherhood has become a trend. Five countries, compared with two only in 1995, are now available for surrogate motherhood, though regulated by strict criteria. Based on the data collected and issues discussed, a detailed guideline may not be generally applicable as each region has its own rules, reflecting differences in the management of ART, especially in the surrogate motherhood. This Committee therefore recommends that ART must be performed by a board certified gynecologist in a certification laboratory and surrogate motherhood must be used only for medical reasons and not be commercialized. The patients of this special group must be treated individually, based on this recommendation and the regulations of their own country/region.

Asia↗

Measurement of the size and orientation of human masseter and medial pterygoid muscles.

To gain a better understanding of biting and chewing performance, the size and orientation of the masseter and medial pterygoid muscles in living humans were studied. Twenty-seven young males having complete dentition, class I dental occlusion and normal muscle and jaw function were examined using magnetic resonance images of the head between the zygomatic arch and hyoid bone. The sections were parallel to the palatal plane, and the thickness was 3 mm without a gap. A computer software program (Medical Dental Image, MDI) was developed to identify and calculate the area of each cross section of the muscle, and the volume of the muscle was then estimated. The axis of the muscle was determined by connecting the centroids of the sections in the lower and upper 1/3 of the whole muscle. The effective muscle cross section area was then calculated by resectioning the muscle perpendicularly to the muscle axis. It was found that the mean masseter muscle volume was around 31 cm3, and that the mean medial pterygoid muscle volume was 11 cm3. Their mean effective cross section areas were around 6.2 cm2 and 3.5 cm2, respectively. The axis of the masseter muscle was more perpendicular to the palatal plane and parallel to the sagittal plane than was the medial pterygoid muscle. The results suggest that the use of magnetic resonance images (MRI) is an effective noninvasive measurement technique for determining the size and orientation of masseter and medial pterygoid muscles. This technique can be employed in future studies on human bite force evaluation and masticatory function.

Adolescent↗

Correlation of cervical vertebra maturation with hand-wrist maturation in children.

The purpose of this study was to evaluate the reliability of cervical vertebra maturation as an indicator of skeletal age during the circumpubertal period. This was determined by correlating cervical vertebra maturation to hand-wrist maturation. The vertebral skeletal age was assessed using lateral cephalometric radiographs according to maturity indicators modified from Lamparski. The hand-wrist skeletal age was evaluated in radiographs with the system developed by Fishman. The sample consisted of 503 subjects (244 boys and 259 girls), aged 8 through 18 years. The Spearman rank correlation coefficients and Wilcoxon sign rank test showed that a statistically significant relationship existed between the two assessments. Both the intra- and inter-judge tests of reliability displayed no significant differences. The results of this study indicate that skeletal age assessment made from the maturational changes of cervical vertebrae were reliable, reproducible and valid.

Adolescent↗

Ductal adenocarcinoma of the prostate with endometrioid features in a 69-year-old man.

We describe a case of ductal adenocarcinoma of the prostate with endometrioid characteristics presenting as painless hematuria and intraurethral tumor. A 69-year-old man had intermittent painless hematuria for 2 months. The serum prostate-specific antigen concentration was elevated (22.0 ng/mL). An enlarged prostate with a necrotic tumor was noted in the right lobe of the prostate on computerized tomography and magnetic resonance imaging studies. A polypoid and worm-like tumor was found within the prostatic urethra near the verumontanum. The tumor had a distinctly papillary configuration with a focal glandular structure on microscopy. Radical prostatectomy was performed and histology of the tumor specimen revealed it to be composed of a closely packed glandular structure lined by single layers of high columnar cells with focal stratification. Frequent papillary projections of glandular epithelium and intraglandular bridging were noted, with a histopathologic appearance similar to endometrioid carcinoma of the uterus. Androgen deprivation therapy was started immediately following surgery. No evidence of recurrence or metastasis was found at follow-up 27 months postoperatively. Distinct features of ductal adenocarcinoma of the prostate include intraurethral papillary tumor close to the verumontanum, urethral obstruction, and easy bleeding of the tumor. Its more aggressive behavior than classical microacinar adenocarcinoma of the prostate makes early recognition of this type of prostatic malignancy important.

Aged↗

Changes in brain cell shape create residual extracellular space volume and explain tortuosity behavior during osmotic challenge.

Diffusion of molecules in brain extracellular space is constrained by two macroscopic parameters, tortuosity factor lambda and volume fraction alpha. Recent studies in brain slices show that when osmolarity is reduced, lambda increases while alpha decreases. In contrast, with increased osmolarity, alpha increases, but lambda attains a plateau. Using homogenization theory and a variety of lattice models, we found that the plateau behavior of lambda can be explained if the shape of brain cells changes nonuniformly during the shrinking or swelling induced by osmotic challenge. The nonuniform cellular shrinkage creates residual extracellular space that temporarily traps diffusing molecules, thus impeding the macroscopic diffusion. The paper also discusses the definition of tortuosity and its independence of the measurement frame of reference.

Animals↗

Expression of alpha 1D subunit mRNA is correlated with L-type Ca2+ channel activity in single neurons of hippocampal "zipper" slices.

L-type voltage-sensitive Ca(2+) channels (L-VSCCs) play an important role in developmental and aging processes, as well as during normal function of brain neurons. Here, we tested a prediction of the hypothesis that membrane density of functional L-VSCCs is regulated by the level of gene expression for its alpha(1D) pore-forming subunit. If so, alpha(1D) mRNA and L-VSCC activity should be positively correlated within individual neurons. Conventional methods of aspiration and/or acute cell dissociation used in prior single-cell studies have generally yielded variable and incomplete recovery of intracellular mRNA. Thus, quantitative relationships between channel function and expression have been difficult to define. In this study, we used the partially dissociated ("zipper") hippocampal slice preparation as a method for collecting a single neuron's mRNA complement. This preparation, developed to expose neuronal somata for recording, also enables the extraction of a neuron with major processes largely intact. Thus, single-cell measures of gene/mRNA expression can be based on approximately the cell's full set of mRNA transcripts. In adult and aged rat hippocampal zipper slices, L-VSCC activity was first recorded in CA1 neurons in cell-attached patch mode. The same neurons were then extracted and collected for semiquantitative reverse transcriptase-PCR analysis of alpha(1D) and calmodulin A (CaM) mRNA content. Across multiple single neurons, a significant, positive correlation was found between the rank orders of L-VSCC activity and of alpha(1D), but not CaM, mRNA expression. Thus, these studies support the possibility that the level of alpha(1D) gene expression regulates the density of functional L-VSCCs.

Animals↗

Concurrent quantification and pharmacokinetic analysis of cefotaxime in rat blood and brain by microdialysis and microbore liquid chromatography.

A simple but effective coupling of microdialysis and microbore liquid chromatograph with UV detection technique was applied to the simultaneous studying of the pharmacokinetics of cefotaxime in both the peripheral compartment and central nervous system. The mobile phase consisted of methanol-100 mM monosodium phosphoric acid (25:75, v/v, pH 5.5) pumped through a C18 microbore column at a flow-rate of 0.05 ml/min. Detection of cefotaxime was set at a UV wavelength of 254 nm. Microdialysis probes were inserted into the jugular vein and striatum of the rat. Following stabilization of microdialysate levels, rats received cefotaxime (20 mg/kg, i.v., n=6) via the femoral vein, and complete concentration versus time profiles for blood and striatum were constructed. The results indicated that cefotaxime rapidly (within 10 min) entered the extracellular fluid of brain striatum following intravenous administration. Noncompartmental pharmacokinetics analysis indicated that the area under the concentration versus time ratio of cefotaxime in rat brain and blood was 6.9%, suggesting appreciable blood-brain barrier penetration. The method was relatively simple, imposed minimal physiological perturbance as it involved no body fluid consumption and sampled in particular protein-unbound drugs, generally believed to be the active fraction.

Animals↗

Effects of Nd:YAG laser irradiation on cultured human gingival fibroblasts.

BACKGROUND AND OBJECTIVE: The Nd:YAG laser has been proposed to apply in minor soft tissue surgery, including various periodontal procedures. However, little information is available regarding the direct effect of Nd:YAG laser on gingival fibroblasts, which play an important role in the early healing processes of periodontal repair. STUDY DESIGN/MATERIALS AND METHODS: Nd:YAG laser irradiation was performed in pulsed mode on human gingival fibroblasts, which was derived from healthy human gingiva by an explant method. The size of laser diode was 400 microm in diameter. The parameters in laser delivery were pulse energy (50-150 mJ), power output (1.0-3.0 W), pulse rate (10-30 pps), and fixed duration of irradiation (10 seconds). The cell cultures were analysed by cytomorphologic examination under phase-contrast and scanning electron microscope. The vitality was also examined with the help of MTT staining. RESULTS: The area of laser damage on cell culture was circular in shape, with diameter beyond the size of laser diode. By scanning electron microscopy, we observed the cellular damage of cultured gingival fibroblasts induced by Nd:YAG laser irradiation, comparable with the progressive increased power settings. The cytomorphologic changes ranged from disappearance of cellular boundary, loss of identifiable cellular nucleus, and finally cell contraction and vacuolization. Significant decrease in cellular vitality (14% approximately 44%) after laser treatment with irradiation distance of nearly contact was noted. However, 2 mm defocusing irradiation with the same power settings did not significantly decrease cellular vitality. CONCLUSION: Our study demonstrated the cell damaging effects of Nd:YAG laser, ranging from degeneratively cytomorphologic change to cell death, on the cultured human gingival fibroblasts. It provided the dentist a chance to understand the potential hazard of laser application in periodontal treatment. If the energy output is enough for the clinical purposes, Nd:YAG laser with lower pulse energy and corresponding pulse rate should be selected to minimize the damage on adjacent soft tissue.

Cell Survival↗