Biomedical subjects

J C Huang

Publications and source records attributed to J C Huang.

At least 73 records · Page 4Linked to original sources

PubMed1994

Dual role of TFIIH in DNA excision repair and in transcription by RNA polymerase II.

The RNA polymerase II general transcription factor TFIIH is composed of several polypeptides. The observation that the largest subunit of TFIIH is the excision-repair protein XPB/ERCC3 (ref. 1), a helicase implicated in the human DNA-repair disorders xeroderma pigmentosum (XP) and Cockayne's syndrome, suggests a functional link between transcription and DNA repair. To understand the connection between these two cellular processes, we have extensively purified and functionally analysed TFIIH. We find that TFIIH has a dual role, being required for basal transcription of class II genes and for participation in DNA-excision repair. TFIIH is shown to complement three different cell extracts deficient in excision repair: XPB/ERCC3, XPC and XPD/ERCC2. The complementation of XPB and XPD is a consequence of ERCC3 and ERCC2 being integral subunits of TFIIH, whereas complementation of XPC is due to an association of this polypeptide with TFIIH. We found that the general transcription factor IIE negatively modulates the helicase activity of TFIIH through a direct interaction between TFIIE and the ERCC3 subunit of TFIIH.

DNA Helicases↗

PubMed1994

Decreased opsin mRNA and immunoreactivity in progressive rod-cone degeneration (prcd): cytochemical studies of early disease and degeneration.

Opsin mRNA level and immunoreactivity were examined by in situ hybridization and immunocytochemistry in normal and progressive rod cone degeneration (prcd)-affected dogs. In situ hybridization used 35S- and/or 3H-labeled bovine opsin cRNA probes; immunocytochemistry used six monoclonal mouse anti-bovine opsin antibodies (MAb1) that are specific to different regions of the N-terminal, loop v-vi and the C-terminal domains. Optimal labeling and histological resolution at the single cell level were achieved with semi-thin sections of DGD wax-embedded tissues; it was possible to correlate the cytochemical observations with the disease staging in topographically defined regions that exhibited different disease severity. In early disease (stages 0-1), opsin mRNA levels and immunoreactivity were normal. During the transition from disease to degeneration (stage 2), however, opsin mRNA was reduced sharply; it then rapidly became undetectable in late stages of degeneration (stages 3, 4). Reduction of immunoreactivity was seen with all the MAbs in stages 2 and 3, but the degree of reduction varied remarkably in different regions of the protein molecule; immunoreactivity was reduced more in the cytoplasmic regions, particularly in the phosphorylation sites and the far end of the C-terminal domain. In contrast, the epitopes of the N-terminal domain that are located in the intradiscal compartment were better preserved. It is noteworthy that, in stages 2 and 3, many rod cells still survived despite the decrease in the mRNA level and immunoreactivity. The results indicate that early disease in prcd-affected rods is not initiated by a reduction of opsin mRNA or the protein quantity. However, opsin expression disappears in early degeneration and before cell death. The differences in immunoreactivity with disease may result either from alterations in the protein structure or configuration, or from selective loss of epitopes located in the cytoplasmic domains of the molecule.

Animals↗

PubMed1994

Ferromagnetic high-permeability alloy alone can provide sufficient low-frequency and eddy-current shieldings for biomagnetic measurements.

A low cost, two-shell ferromagnetic shielded room large enough for a person to enter and prepare experiments was constructed for biomagnetic measurements. No aluminium or copper shell was used for eddy-current shielding. A high-permeability nickel-iron-molybdenum alloy (1.57 mm thick) was used for both ferromagnetic and eddy-current shieldings. The shielding factor was 60 dB at dc and 40 dB between 0.1 Hz and about 3 Hz. The eddy-current shielding due to the alloy alone provided a shielding factor of 55 dB at 30 Hz and 66 dB at 60 Hz. The shielding factor was sufficiently high in all the frequency range for biomagnetic measurements with a first-order superconducting gradiometer.

Alloys↗

PubMed1994

Analysis of spermatozoa from the proximal vas deferens of vasectomized men.

This study assessed the condition of spermatozoa from the proximal vas deferens of men after vasectomy. The fluids of both proximal vas deferens were collected from 67 vasectomized men by cannulating the vas deferens at the time of vasectomy reversal. Selected sperm parameters were analysed after incubation of the spermatozoa for 30 min at 37 degrees C. Sperm concentration in the proximal vas from vasectomized men (16,312 +/- 21,496 million per ml, geometric mean: 7948 +/- 398 million per ml) was significantly higher than that of fertile men and was maintained at a constant level independent of the duration of vas obstruction. The means of sperm motility (36.2 +/- 26.2%), spermatozoa with normal morphology (50.7 +/- 21.7%), sperm viability (53.0 +/- 25.3%) and hypo-osmotic swelling test (HOS-test, 53.9 +/- 21.7%) were statistically lower than the respective values for normal fertile men. There was no significant correlation between the duration of vas obstruction and the above semen parameters. In 46.4% of vas fluids all spermatozoa were immotile and this condition was more common after 3 years of vasectomy. Immotile spermatozoa in the proximal vas fluids at the time of vasectomy reversal may be an important factor for predicting semen quality and fertilizing ability after vasovasostomy. There were no significant differences in the results of sperm-cervical mucus penetration test (CMPT) between spermatozoa from vasectomized and fertile men. Antisperm antibodies on the surface of spermatozoa from the vas of vasectomized men were determined by the immunobead test (IBT; 78.6% for IgG, 32.1% for IgA) and sperm cervical mucus contact test (SCMC, 36.4%).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1994

Origin of the apparent tissue conductivity in the molecular and granular layers of the in vitro turtle cerebellum and the interpretation of current source-density analysis.

1. We determined the origin of the apparent tissue conductivity (sigma 2) of the turtle cerebellum in vitro. 2. Application of a current with a known current density (J) along the longitudinal axis of a conductivity cell produced an electric field in the cerebellum suspended in the cell. The measured electric field (E) perpendicular to the cerebellar surface indicated a significant inhomogeneity in sigma a (= J/E) with a major discontinuity between the molecular layer (0.25 +/- 0.05 S/m, mean +/- SD) and granular layers (0.15 +/- 0.03 S/m) (n = 39). 3. This inhomogeneity was more pronounced after anoxic depolarization. The value of sigma a decreased to 0.11 +/- 0.03 and 0.040 +/- 0.008 S/m in the molecular and granular layers, respectively. The ratio of sigma a S in the two layers increased from 1.67 in the normoxic condition to 2.75 after anoxic depolarization. 4. This difference in sigma a across the two layers was present within the range of frequencies (DC to 10 kHz) studied where the phase of sigma a was small (less than +/- 2 degrees) and therefore sigma a was ohmic. 5. The inhomogeneity in sigma a was in part due to an inhomogeneity in the extracellular conductivity (sigma e) as determined from the extracellular diffusion of ionophoresed tetramethylammonium. Like sigma a, the value of sigma e was also higher in the molecular layer (0.165 S/m) than in the granular layer (0.097 S/m). The inhomogeneity in sigma e was due to a smaller tortuosity and a larger extracellular volume fraction in the molecular layer compared with the granular layer. 6. sigma a was, however, consistently higher, by approximately 50%, than sigma e. A core conductor model of the cerebellum indicated that these discrepancies between sigma a and sigma e were attributable to additional conductivity produced by a passage of the longitudinal applied current through the intracellular space of Purkinje cells and ependymal glial cells, with the glial compartment playing the dominant role. Cells with a long process and a short space constant such as the ependymal glia evidently enhance the effective "extracellular" conductivity by serving as intracellular conduits for the applied current. The result implies that the effective sigma e may be larger than sigma e for neuronally generated currents in the turtle cerebellum because the space constant for Purkinje cells is several times greater than that for the ependymal glia and consequently Purkinje cell-generated currents travel over a long distance relative to the space constant of glial cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1994

Quantitative analysis of epidermal growth factor receptor gene expression in endometriosis.

The molecular mechanisms by which endometriosis persists in locations outside the uterus are unclear. Recently, the epidermal growth factor receptor (EGF-R) has been postulated to have a role in the disease process of endometriosis. To explore this, we determined the levels of EGF-R protein and messenger ribonucleic acid (mRNA) expression in endometriotic tissues and compared the levels to that of eutopic endometrium. Using rabbit anti-EGF-R antibody, we found more intense immunohistochemical staining for EGF-R in glandular cells than in stromal cells of both endometriomas and endometriotic implants. No difference in staining intensity was noted between endometriotic tissues and eutopic endometrium. A ribonuclease protection assay was used to determine mRNA levels for EGF-R. PhosphoImager analysis revealed the following levels of mRNA for EGF-R; eutopic endometrium, 1.00 +/- 0.27 (arbitrary units; mean +/- SEM; n = 6 patients); cyst walls of endometriomas, 0.21 +/- 0.12 (n = 10 patients); endometriotic implants, 0.29 +/- 0.13 (n = 9 patients); and pelvic adhesions, 0.03 +/- 0.03 (n = 5 patients). Endometriotic tissues had significantly less mRNA for EGF-R than eutopic endometrium (P < 0.05, by Newman-Keuls test). Our findings support the hypothesis that EGF-R may be associated with the disease process of endometriosis.

Endometriosis↗

PubMed1994

Spontaneous rupture of the liver associated with pregnancy: a report of two cases.

Spontaneous rupture of the liver associated with pregnancy is a rare and grave complication, usually occurring in preeclampsia or eclampsia. Two cases of ruptured subcapsular hematoma of the right liver during pregnancy are reported. The first case was a 19-year-old woman who had suffered from epigastralgia and absent fetal heart beat in the 32nd week of gestation. The second case was a 31-year-old female who complained of nausea and right upper quadrant pain in the 35th week of pregnancy. Both had preeclampsia, and developed shock with disseminated intravascular coagulation soon after admission. Both received surgery and were found to have ruptured hematoma over the right liver. Finally, the first patient died of renal failure, but the second survived because preoperative diagnosis had been exact. Greater suspicion, then awareness of diagnosis can lead to better timing of surgery and an improved prognosis for mother and child.

Adult↗

PubMed1993

A chemiluminescent substrate for the detection of phosphatidylinositol-specific phospholipase C.

A water-soluble, chemiluminescent substrate for sensitive detection of bacterial phosphatidylinositol-specific phospholipase C (PI-PLC) activity was synthesized. This was achieved by replacing the diacylglycerol moiety found in the natural substrate, phosphatidylinositol, by a dioxetane-containing moiety, resulting in the synthetic substrate (+/-)-3-(4-methoxyspiro[1,2-dioxetane-3,2'- tricyclo[3.3.1.1(3,7]decan]-4-yl)phenyl myo-inositol-1-O-hydrogen phosphate (LUMI-PI). PI-PLC-catalyzed cleavage of the phosphodiester bond of LUMI-PI results in the formation of a chemiluminescent precursor, the labile anion AMP-D, which decays with emission of light. An assay procedure was developed using 96-well microtiter plates with detection of the chemiluminescence on autoradiography film. With this procedure the detection limit of purified phospholipase C was about 10 pg. Furthermore, PI-PLC activity was readily detected in situ using small quantities (< 1 microliters) of liquid cultures of PI-PLC-producing bacterial strains.

Adamantane↗

PubMed1993

Inhibition of beta-adrenoceptor agonist relaxation of airway smooth muscle by Ca(2+)-activated K+ channel blockers.

In isolated guinea pig trachea contracted by 0.5 mM acetylcholine, the cumulative relaxant concentration-response curves to the beta 2-adrenoceptor agonist, salbutamol, were shifted to the right by depolarizing concentrations of KCl, as well as by charybdotoxin, iberiotoxin and tetraethylammonium ion, which are antagonists of the high-conductance Ca(2+)-activated K+ channel. The shifts produced by KCl (40 mM), charybdotoxin (100 nM), iberiotoxin (50 nM), and tetraethylammonium ion (2 mM) were approximately 230-fold, 10-fold, 78-fold, and 8-fold, respectively. The blockade of beta 2-adrenoceptor agonist-induced relaxation by these agents was totally reversed by 0.3 microM nifedipine. Similar reversal was obtained with either 100 microM CdCl2, or low Ca2+ (50 microM) Krebs medium. These data suggest that charybdotoxin, iberiotoxin and tetraethylammonium ion, like KCl, cause membrane depolarization which in turn activates voltage-dependent Ca2+ channels. The influx of Ca2+ via these channels provides an additional mode to that of release of intracellular Ca2+ evoked by acetylcholine for maintaining cell Ca2+ concentration at a high level. This is apparently sufficient to account functionally for the blockade of beta 2-adrenoceptor agonist-induced relaxation. In view of this interpretation regarding the action of Ca(2+)-activated K+ channel antagonists, earlier proposals ascribing the relaxant effect of beta 2-adrenoceptor agonists strictly to activation of these channels must be reevaluated.

Acetylcholine↗

PubMed1993

Survival of cone responses in postmortem human retina.

A cone-mediated electroretinogram was recordable from human retinas 17-18 hours postmortem, after regeneration of visual pigments by application of 11-cis and 9-cis retinal. The cone was separated from the rod component by stimulation with flickering red light. In addition to these neuronal responses, a P2 and a slow P3 were present.

Adolescent↗

PubMed1993

Diethylene glycol distearate (DGD): a versatile embedding medium for retinal cytochemistry.

Embedment in diethylene glycol distearate (DGD) was shown to be highly desirable and versatile for retinal cytochemical studies, including in situ hybridization, immuno- and lectin cytochemistry. This method allows for preservation of fine tissue detail as well as good reaction sensitivity. It appears to be more suitable than most other methods currently used for light microscopic retinal cytochemistry.

Animals↗

PubMed1993

Congenital absence of vas deferens treated by artificial spermatocele constructed from the tunica vaginalis; report of six cases.

Six patients with congenital absence of the vas deferens were treated by artificial spermatoceles (ASCs) in an attempt to collect sperm for subsequent artificial insemination (AI). The ASC was constructed from a flap of tunica vaginalis from the patient. Sperm were aspirated from the ASC each month; there was wide variation in the concentration of sperm recovered [33.0 +/- 48.2 x 10(6) mL-1 (range 0.1-200)] and motility and normal morphology of the sperm were 19.0 +/- 17.9% and 48.8 +/- 23.4% respectively. Although a total of 14 cycles of AI was performed using the aspirated sperm, pregnancy did not result. Both poor sperm recovery and poor motility contributed to this failure. The condition of the epididymis observed during scrotum examination is reported. It is suggested that in some patients with congenital absence of vas deferens, the paired Wolffian ducts may differ from each other in their development.

Adult↗

PubMed1993

Temporal variation in bacterial disease frequency: molecular population genetic analysis of scarlet fever epidemics in Ottawa and in eastern Germany.

In an effort to understand the molecular genetic basis of temporal variation in frequency and severity of bacterial disease, genetic relationships among strains of Streptococcus pyogenes that caused scarlet fever epidemics in Canada in the early 1940s and in eastern Germany in the 1960s to 1980s were studied. Application of multilocus enzyme electrophoresis and comparative sequencing of the gene (speA) encoding streptococcal pyrogenic exotoxin A (scarlet fever toxin) revealed that new waves of scarlet fever are associated with an increase in frequency of S. pyogenes clones carrying variant speA alleles. This finding suggests that the occurrence of new scarlet fever epidemics can be predicted by comprehensive monitoring of the frequency of S. pyogenes clones with variant toxin alleles.

Alleles↗

PubMed1993

Cytomegalovirus infection of the cervix detected by cytology and histology: a report of five cases.

We present four cases of cytomegalovirus (CMV) infection detected by cervical cytology of asymptomatic young women. One patient who had undergone bone marrow transplantation was immunosuppressed, but no factors predisposing to CMV infection were identified in the other three patients. A cervical biopsy specimen from a fifth patient, who was also asymptomatic, demonstrated the locus of CMV infection to be in endocervical gland cells. Immunocytochemical and polymerase chain reaction studies on this biopsy specimen confirmed that the histological changes were caused by CMV. The finding of CMV-infected cells in cervical cytological or biopsy specimens is a rarity. Our observations in asymptomatic, non-immunocompromised women suggest their presence is an incidental finding, unlikely to have any clinical significance.

Adult↗

PubMed1993

[Effects of scoparone on hemodynamics in anesthetized rabbits].

The effects of scoparone (Sco), tetramethylpyrazine (TMP), and papaverine (Pap) on hemodynamics were compared in anesthetized rabbits. Three dosage levels (1.25, 2.5, 5 mg.kg-1, iv) were used for each agent. Sco lowered the blood pressure (BP), increased the cerebral blood flow (CBF) dose-dependently, but did not show significant influence on the femoral artery blood flow (FBF), the maximal decrease of BP, and the maximal increases of CBF being 15% and 20% of the control, respectively. In contrast, TMP raised the BP as well as increased the CBF and FBF simultaneously. Pap, however, lowered the BP, slightly increased the CBF and decreased the FBF only at large doses. The results suggested that Sco may cause a selective dilatation of cerebral blood vessels.

Animals↗

PubMed1993

[Effects of 6,7-dimethoxycoumarin on left ventricular function and hemodynamics in anesthetized cats].

Scoparone (6,7-dimethoxycoumarin 1.25, 2.5, 5 mg.kg-1) influenced the cardiohemodynamic functions in anesthetized cats. After i.v. injection the LVP-dP/dt loops and blood pressure were decreased. Both the contractile and relaxant functions of the left ventricle were inhibited. These were demonstrated by the reduction of dP/dtmax, Vmax, Vce-dp40, LVSP, prolongation of LVET, diminution of -dP/dtmax, and extension of the constant time of isometric diastolic pressure decay (T value). The preload of the left ventricle, LVEDF, was raised and the afterload (MAP, TPB) was decreased: The systolic volume, systolic index, cardiac output, and cardiac index were decreased slightly. The parameters TTI and TTI x HR, which reflect the myocardial oxygen consumption, were reduced. Furthermore, the heart rate was decreased at 5 mg.kg-1. The results indicate that scoparone could suppress the myocardial activity.

Animals↗

PubMed1992

Human nucleotide excision nuclease removes thymine dimers from DNA by incising the 22nd phosphodiester bond 5' and the 6th phosphodiester bond 3' to the photodimer.

By using a human cell-free system capable of nucleotide excision repair, a synthetic substrate consisting of a plasmid containing four thymidine dimers at unique locations, and deoxyribonucleoside 5'-[alpha-thio]triphosphates for repair synthesis, we obtained DNA fragments containing repair patches with phosphorothioate linkages. Based on the resistance of these linkages to digestion by exonuclease III and their sensitivity to cleavage by I2, we were able to delineate the borders of the repair patch to single-nucleotide resolution and found an asymmetric patch with sharp boundaries. That the repair patch was produced by filling in a gap generated by an excision nuclease and not by nick-translation was confirmed by the finding that the thymidine dimer was released in a 27- to 29-nucleotide oligomer.

Base Sequence↗

PubMed1992

Effects of dietary chloride and magnesium on the incidence of tibial dyschondroplasia in chickens fed on Chinese practical diets.

1. The effects of varying the concentrations of dietary magnesium and chloride on the performance, incidence of tibial dyschondroplasia and tissue element contents of broilers fed on Chinese practical maize soya-bean meal diets was assessed. 2. Two concentrations of dietary chloride (1.5 and 3.5 g/kg) and 3 of dietary magnesium (2.0, 4.0 and 6.0 g/kg) were used to form 6 dietary treatments for a 21-d experimental period. 3. Results from this study indicated that the dietary ingredients of maize and soya-bean meals produced in the northeast of China may predispose or influence the susceptibility of chickens to tibial dyschondroplasia. 4. High dietary chloride induces a high incidence of tibial dyschondroplasia (P less than 0.05) which could be ameliorated by dietary magnesium supplementation. 5. Dietary magnesium plays an anti-rachitic role by improving the deposition of copper and zinc in the bone tissue. 6. Supplementation with magnesium to 4.0 g/kg is recommended for broiler diets in northeast China.

Animal Feed↗