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Biomedical subjects

K C Chang

Publications and source records attributed to K C Chang.

At least 19 recordsLinked to original sources

Transformation of a novel direct-repeat repressor element into a promoter and enhancer by multimerisation.

Studies on the regulation of interferon (IFN) responsive genes have mainly been centred on the highly conserved IFN stimulated responsive elements (ISREs) which can mediate type I and II IFN inducibility. To date little is known about other functional cis-acting regulatory motifs in IFN responsive genes. We report here on the identification of a repressor element in the human MxA gene defined to a 19 base pair (bp) region which houses a 9 bp direct repeat. DNA-specific protein binding on this element is not affected by IFN treatment and is distinct from ISRE binding proteins. Remarkably, contrary to expectations, when the repressor element is multimerised and spliced, in either orientation, to a reporter gene it behaves like a functional, constitutive promoter. Positioning the multimerised element in front of the SV40 enhancerless promoter also led to enhanced expression. The same protein(s) seem to bind to both the single repressor element and its multimerised form. This discovery of phenotypic reversal on a repressor element via multimerisation may have important implications in vivo.

Base Sequence

Pharmacological evaluation of GS-389, a novel tetrahydroisoquinoline analog related to higenamine, on vascular smooth muscle.

The effects of GS-389, a novel tetrahydroisoquinoline analog, on isolated rat and mouse thoracic aorta rings, were investigated. Both GS-389 and papaverine induced endothelium-independent, concentration-dependent relaxations of the rat and mouse aortae precontracted with phenylephrine (PE). The GS-389-induced inhibition of the contractile response to PE was noncompetitive. The initial phasic contraction to PE elicited in Ca(2+)-free media was also attenuated by pretreatment with GS-389, indicating that GS-389 may interfere with the release of intracellular Ca2+ and/or the effects of intracellular Ca2+ release. GS-389 potentiated the vasodilatory effects of isoproterenol and sodium nitroprusside in rat and mouse aortae. GS-389 significantly increased cGMP levels in the rat aorta and inhibited cGMP phosphodiesterase from the rabbit brain. Methylene blue, but not propranolol, inhibited the vasodilatory effect of GS-389. These results suggest that the vasorelaxant effect of GS-389 may be due, at least in part, to inhibition of cGMP metabolism.

Alkaloids

Dibutyryl cyclic AMP and forskolin inhibit phosphatidylinositol hydrolysis, Ca2+ influx and contraction in vascular smooth muscle.

Dibutyryl cyclic AMP and forskolin inhibited the contraction induced by norepinephrine (NE) more strongly than the high K(+)-induced contraction in isolated rat aorta. These inhibitors inhibited the 45Ca2+ influx stimulated by NE but not that by high K+, and they inhibited NE-induced inositol monophosphate accumulation. These results suggest that cAMP inhibits NE-induced contraction, at least partly, by inhibiting the alpha-adrenoceptor-mediated signal transduction and high K(+)-induced contraction by decreasing Ca2+ sensitivity but not Ca2+ influx.

Animals

[Multiple leiomyosarcomas of extremities].

Leiomyosarcoma is an uncommon tumor that rarely occurred in extremity. Multiple leiomyosarcomas in different extremities is even rarer. We report a 90 year-old male veteran who was found to have four leiomyosarcomas, one in right arm and the other three in right thigh. The patient sustained gunshot injuries over right thigh and right arm about 60 years ago in a war with severe wound sepsis due to open communited fracture of the femur. These tumors happened to distribute in the vicinity of the old scars. It was speculated that these multicentric tumors were due to decreased immune surveillance caused by aging and induced by the previous trauma. The literatures were reviewed and the diagnostic methods, surgical treatment and the adjuvant therapy were discussed.

Aged

Strong expression of foreign genes following direct injection into fish muscle.

We report here for the first time direct injection of genes into fish muscle in vivo. Plasmids used contain either SV40 early promoter, rabbit beta-cardiac myosin heavy chain promoter, human MxA promoter or an artificial promoter, fused to a chloramphenicol acetyltransferase (CAT) or beta-galactosidase reporter gene. CAT assays revealed that most gene constructs were highly expressed. Histochemical analysis showed that beta-galactosidase was strongly expressed at the site of injection within muscle fibres. This method provides an excellent system for testing expression of gene constructs, including those of mammalian origin, in fish muscle in vivo and has the potential for fish vaccination.

Age Factors

Molecular and functional analysis of the virus- and interferon-inducible human MxA promoter.

The virus- and interferon-inducible human MxA (IFI-78k) gene is a homologue of the murine influenza resistance gene Mx1. Three overlapping human cosmid clones covering most of the gene including its promoter region were isolated. Sequencing the 5' MxA cDNA derived by RT-PCR (reverse transcriptase-polymerase chain reaction) confirmed the most 5' putative transcriptional start site. The MxA promoter does not contain a TATA or CCAAT box but has three Interferon Stimulated Response Element (ISRE) motifs. Strong induction with type I interferons was demonstrated with a fragment containing only two ISREs in human L132 cells. This induced expression was not adversely affected by 2-aminopurine. However, the promoter showed constitutive expression in transiently or stably transfected murine LM cells.

Animals

Variations in microbial indicator densities in beach waters and health-related assessment of bathing water quality.

Daily and hourly variations in microbial indicators densities in the beach-waters of Hong Kong have been described. The levels of Escherichia coli at a number of beaches was observed to be influenced by tide, and for staphylococci, by bather numbers. The tidal influence was most obvious during spring tides; and for the effect of bathers, during neap tides. Both organisms are present in high densities in external sources of faecal pollution of bathing beaches, with the average staphylococci to E. coli ratios being 0.04-3. Staphylococci may serve as an indicator of bather density and the risk of cross-infection amongst bathers (rather than as another indicator of faecal contamination) when the average staphylococci to E. coli ratio for a bathing beach is considerably higher than 3. The variability of microbial indicator densities means the routine sampling of bathing beaches should be carried out on weekend days with maximum numbers of swimmers exposed to the water, and spread throughout the bathing season.

Colony Count, Microbial

Inhibition of phorbol ester-induced contraction by calmodulin antagonists in rat aorta.

The purpose of the present study was to investigate the relative roles of protein kinase C (PKC) and myosin light chain kinase (MLCK) in phorbol ester-induced contraction of vascular smooth muscle through the use of PKC and calmodulin antagonists. Prior exposure to PKC antagonists staurosporine (0.03 microM) and H-7 (10 microM) had relatively little effect on contractions to phorbol 12-myristate 13-acetate (PMA), while contractions to norepinephrine and KCl were greatly inhibited. Prior exposure to the calmodulin antagonists calmidazolium (3 and 10 microM) and W-7 (10 microM) inhibited contractions to PMA in the presence and absence of extracellular Ca2+, while contractions to norepinephrine and KCl remained relatively unaffected. Calmidazolium and W-7 were relatively weak relaxants when applied during the PMA contraction, and the magnitudes of relaxation were similar to those observed in norepinephrine- and KCl-contracted tissues. Calmidazolium partially inhibited the PMA-induced translocation of PKC. These results suggest that 1) the calmodulin antagonists inhibit the development of PMA-induced contraction, at least in part, through inhibition of PKC translocation; 2) the mechanisms of phorbol ester- and agonist-induced translocation of PKC are distinct; 3) the potencies and inhibitory mechanisms of these agents depend on whether the agents are added before or during the contraction; and 4) the selectivity of these agents, as evaluated in enzyme preparations, may not be consistent with their cellular actions.

Animals

Vascular factors in isovolumic systemic and pulmonary circuit.

Experiments were conducted in 12 pentobarbital-anesthetized dogs with sinus denervation and vagotomy. The chest was opened, and the heart was replaced by a roller pump with two perfusion lines. The systemic and pulmonary circulations (SC and PC) were perfused with a constant and adjustable flow (Q). Venous outflows were directly driven by pumps without passing through a reservoir. In each closed circuit, the total blood volume remained constant because inflow and outflow were simultaneously and equally altered. In both SC and PC, arterial pressure (Pa), i.e., systemic arterial and pulmonary arterial pressures (SAP and PAP), was a positive function of Q, and venous pressure (Pv), i.e., right atrial and left atrial pressures (RAP and LAP), was a negative function of Q. The first series of experiments involved three equal step reductions in Q from baseline to zero flow. The venous-to-arterial compliance ratio (Cv/Ca) was calculated from delta Pa/delta Pv and vascular resistance (VR) from (Pa - Pv)/Q. The values of Cv/Ca in SC increased from 9.3 +/- 0.4 to 14.5 +/- 1.1 and 21.9 +/- 1.4 (P less than 0.001) in the three-step Q reduction. VR in the SC was not significantly dependent on Q. In the PC, the Cv/Ca was approximately 2.0 and VR was 0.16 mmHg.ml-1.min.kg, both values being independent of Q. Multiple-step reduction in Q for regression equations was carried out in 8 of the 12 dogs. We found that only the SAP was a linear function of Q (ml.min-1.kg-1): SAP = 18.458 + 0.953Q.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Assessment of threshold and saturation pressure in the baroreflex function curve: a new mathematical analysis.

The baroreflex function has been assessed with logistic function analysis, a mathematical model suitable for function curves of sigmoid shape. The nonlinear (sigmoid) relationship between carotid sinus pressure (CSP) and systemic arterial pressure (SAP) reflects threshold and saturation of the SAP responses to CSP changes. The threshold and saturation pressures (TP and SP) are often determined by gross inspection from the experimental recordings. Objective and accurate determination of TP and SP may be achieved by mathematical analysis. Kent et al. (Cardiology, Vol. 57, pp. 295-310, 1972) provided equations that estimated SP and TP to be +/- 1.317/k from the midrange CSP (k, the slope coefficient of a curve). Tan et al. (Circ. Res., Vol. 65, pp. 63-70, 1989) recently used an equation to calculate TP based on the consideration that it is about 95% of the maximal SAP. In this report, we elaborated new equations for the estimation of SP and TP, which approximate midrange pressure +/- 2/k. The accuracy of these new equations compared to other equations was tested using various sets of simulation data. In addition, experiments were conducted in anesthetized dogs with isolated carotid sinus. The open-loop CSP-SAP curves were obtained following various holding pressure (HPs) to demonstrate the phenomenon of baroreflex acute resetting. The effects of using different equations on the values of TP, SP, and delta TP/delta HP (extent of resetting) were analyzed. Both simulation and experimental data revealed that the equations of Kent et al. gave rise to values of TP and SP far different from the realistic values. The values of TP calculated by equation of Tan et al. were dependent on the maximal SAP and the slope of the function curve. The TP and SP (+/- 2/k from the midrange pressure of the sigmoid curve) obtained by our new equations were not significantly affected by the maximal SAP and the curve slope. The mathematical analysis may be particularly useful for comparison among various baroreflex curves with different maximal SAP and/or curve slope.

Animals

Effect of loading conditions on peak aortic flow velocity and its maximal acceleration.

Aortic flow measurement with Doppler echocardiography has become a non-invasive technique in clinical practice. In the present animal study, we evaluated the flow-derived parameters such as peak velocity (PV) and its maximal acceleration (MA) as indices of ventricular contractility independent of the loading status. Eight pentobarbital-anesthetized cats were maintained with artificial ventilation. The chest was opened to place an electromagnetic flow probe around the ascending aorta for recording pulsatile aortic flow. PV and MA were measured from the flow tracing and on-line electronic differentiation. Intravenous infusions of dobutamine (DT), angiotensin II (AII) and dextran (DN) were used to alter the cardiac inotropism, afterload and preload, respectively. At a steady state (approximately 5 min after infusion), DT increased the PV from 56 +/- 9 to 78 +/- 14 cm/sec (p less than 0.05) and MA from 1302 +/- 108 to 1699 +/- 117 cm/sec2 (p less than 0.05). In response to AII infusion, PV was slightly reduced (60 +/- 7 to 55 +/- 6 cm/sec, p less than 0.05) while MA was also reduced mildly but significantly (1219 +/- 109 to 1099 +/- 109 cm/sec2, p less than 0.05). Dextran infusion produced a marked increase in PV (48 +/- 7 to 82 +/- 13 cm/sec, p less than 0.05) while the increase was slightly less for MA (1089 +/- 95 to 1604 +/- 109 cm/sec2). The results indicated that inotropic stimulation markedly increased both PV and MA. PV and MA responded slightly but significantly to afterload alterations. (8.3% vs 9.8%, respectively). Both PV and MA increased markedly to the preload increment.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II

[Versatility of rectus abdominis muscle or myocutaneous flap--report of seven cases].

The rectus abdominis muscle, due to its strategic location in the abdomen with its dual blood supply and wide arc of transposition, is ideally suited to repair defects over the chest, abdomen, groin and perineum. The muscle is generally based on the superior epigastric artery, deep inferior epigastric artery and their associated capillary communications. From September 1989 to September 1990, at Taichung Veterans General Hospital, seven cases were presented utilizing unilateral rectus abdominis muscle or myocutaneous flap for local or free tissue transfer. The recipient areas included abdomen, groin, perineum, lower leg and a penis reconstruction. Of all flaps that survived, one developed minor skin necrosis causing urethral orifice stricture and urethrocutaneous fistula that later required repair. All cases achieved the surgical goals. There was no donor site infection, nor abdominal wall herniation but persisted wound pain in 2 cases was noted during the follow-up period. All patients resumed daily activity without limitation about six weeks postoperatively. The large skin territory with abundant soft tissue offers versatile flap design. The dissection is simple, speedy and straight foreward. The deep inferior epigastric vessels are long and their calibers are suitable for vascular anastomosis. Usually, the donor sites are primary closed. We recommend this flap as one to be considered in reconstruction planning and as a donor material for free tissue transfer.

Adolescent

Studies in the in vivo expression of the influenza resistance gene Mx by in-situ hybridisation.

The inbred laboratory mouse strain A2G carries a functional, interferon type 1 inducible gene, Mx which upon expression confers specific resistance to an otherwise lethal dose of influenza virus. We investigated in vivo Mx gene expression by performing Northern hybridisation and in-situ hybridisation on A2G (Mx+) and (CBA/J x C57)F1 (Mx-) mice that were induced either with human, natural interferon; human, recombinant interferon or double stranded poly(I):(C). All 3 inducers were able to stimulate Mx expression in all organs examined in the A2G strain. However, contrary to previous reports, Mx expression was confined to a small number of cell types; the main contributor was most probably mononuclear cells. Specialised cells such as hepatocyte, nephron, ovarian follicle and seminiferous tubules did not show detectable Mx level. There was also constitutive Mx expression in the epithelia of uterus and duodenum which suggested direct gene activation independent of blood-bourne interferon.

Animals

Inositol uptake in rat aorta.

The purpose of this study was to investigate the mechanism of inositol uptake into rat thoracic aorta. 3H-inositol uptake into deendothelialized aorta was linear for at least 2 h and was composed of both a saturable, Na(+)-dependent, and a nonsaturable, Na(+)-independent component. The Na(+)-dependent component of inositol uptake had a Km of 50 microM and a Vmax of 289 pmol/mg prot/h. Exposure to LiCl, ouabain, or Ca2(+)-free Krebs-Ringer bicarbonate solution inhibited uptake. Metabolic poisoning with dinitrophenol, as well as incubation with phloretin, an inhibitor of carrier-mediated hexose transport, also inhibited uptake. Exposure to norepinephrine decreased inositol uptake, while phorbol myristate acetate was without effect. Isobutylmethylxanthine significantly increased inositol uptake, while the increased uptake due to dibutyryl cyclic AMP and forskolin were not statistically significant. Sodium nitroprusside, an activator of guanylate cyclase, and 8-bromo cyclic GMP, were without effect on uptake, as was methylene blue, an inhibitor of guanylate cyclase. Inositol uptake into the aorta was increased when the endothelium was allowed to remain intact, although this effect was likely due to uptake into both the endothelial and smooth muscle cells. These results suggest that the uptake of inositol into vascular smooth muscle is: (1) dependent upon an inward Na(+)-gradient; (2) carrier mediated, and (3) inhibited by alpha 1 adrenoceptor agonists.

1-Methyl-3-isobutylxanthine

Urinary gonadotropin fragment, a new tumor marker. III. Use in cervical and vulvar cancers.

The efficacy of urinary gonadotropin fragment (UGF) and squamous cell carcinoma antigen (SCC) measurements was examined in the management of cervical and vulvar cancers. Of women with benign gynecologic disease (n = 89) 7%, of women with cervical or vulvar intraepithelial neoplasia (n = 25) 8%, and of those with cervical or vulvar malignancies (n = 60) 47% had elevated UGF levels (greater than 3 fmol/ml). SCC at a cutoff of 2.5 ng/ml had a similar sensitivity, 43%, for the same group of cervical and vulvar malignancies. The populations recognized by SCC and UGF, however, only partially overlapped, so that together UGF and SCC were elevated in 62% of women with malignancies. The sensitivity of both markers was stage dependent, so that UGF and SCC detected 26 and 22%, respectively, of early (stage I or II), and 67 and 40%, respectively, of advanced (stage II or IV) cancers. We monitored the progress of 21 women undergoing therapy for cancer with UGF and SCC measurements. Of the 21, 13 (62%) had true-positive UGF levels and 11 (52%) had true-positive SCC levels when cancer was initially detected. The levels of UGF accurately reflected changing clinical observations in 11 of 21 (52%); those of SCC, in 6 of 21 (29%); and levels of either, in 14 of 21 (67%) cases. Recurrences occurred in 7 of the 21 cases. Rising SCC levels predicted (at an earlier clinic visit) the recurrence in 4 of the 7, and rising UGF levels in 5 of the 7 (includes the 4 detected by SCC) patients. While these numbers for UGF and SCC sensitivity are not ideal, until other markers become available, they are seemingly the best achievable. It is suggested that both UGF and SCC be used to monitor therapy and to detect recurrences of cervical and vulvar cancers.

Antigens, Neoplasm

Health effects of beach water pollution in Hong Kong.

Prospective epidemiological studies of beach water pollution were conducted in Hong Kong in the summers of 1986 and 1987. For the main study in 1987, a total of 18741 usable responses were obtained from beachgoers on nine beaches at weekends. The study indicated the overall perceived symptom rates for gastrointestinal, ear, eye, skin, respiratory, fever and total illness were significantly higher for swimmers than non-swimmers; and the swimming-associated symptom rates for gastrointestinal, skin, respiratory and total illness were higher at 'barely acceptable' beaches than at 'relatively unpolluted' ones. Escherichia coli was found to be the best indicator of the health effects associated with swimming in the beaches of Hong Kong. It showed the highest correlation with combined swimming-associated gastroenteritis and skin symptom rates when compared with other microbial indicators. A linear relationship between E. coli and the combined symptom rates was established. Staphylococci were correlated with ear, respiratory and total illness, but could not be used for predicting swimming-associated health risks. They should be used to complement E. coli. The setting of health-related bathing-water quality standards based on such a study is discussed.

Adolescent

Effects of nifedipine on systemic hydraulic vascular load in patients with hypertension.

STUDY OBJECTIVE: The aim of the study was (1) to determine the difference in aortic input impedance and derived parameters between hypertensives and normotensives; and (2) to assess the acute effects of nifedipine on the aortic impedance, compliance, and resistance in patients with hypertension. DESIGN: A high fidelity multisensor catheter (Millar) was used to obtain the aortic pressure and flow signals for impedance analysis. The acute effects of nifedipine on the impedance parameters were evaluated at steady state before and after (10-30 min) a sublingual dose of 10 mg. PATIENTS: The patients included seven normotensive (mean blood pressure, 97 mm Hg) and nine hypertensive (mean blood pressure, 135 mm Hg), age matched, ethnic Chinese. Patients with clinical evidence of heart failure and valvular or congenital heart diseases were excluded. MEASUREMENTS AND MAIN RESULTS: Pulsatile aortic flow and pressure were measured by Millar catheter inserted into the ascending aorta. Cross sectional area of aorta was estimated by echocardiograms. Cardiac output was determined by Fick principle with an oximeter. These data were subjected to Fourier analysis for impedance spectra. In comparison with normotensives, hypertensives had increased peripheral vascular resistance R, at 2751(705) v 1651(363) dyne.s.cm-5; increased characteristic impedance Zc, at 193(64) v 122(27) dyne.s.cm-5; and increased first zero crossing frequency of impedance phase angle fo, at 4.8(0.9) v 3.4(0.7) Hz. Arterial compliances corresponding to peak systolic pressure Cs were lower, at 0.32(0.19) v 0.90(0.32) ml.mm Hg-1, as was mean pressure Cm, at 0.55(0.25) v 1.24(0.38) ml.mm Hg-1, and end diastolic pressure Cd, at 0.83(0.29) v 1.65(0.44) ml.mm Hg-1. Although the values of external ventricular hydraulic power were higher in hypertensive subjects, the difference was not statistically significant. Nifedipine administration in 7/9 hypertensives significantly reduced R, from 2806(721) to 2433(664) dyne.s.cm-5; mean aortic pressure Pm, from 138(22) to 112(12) mm Hg; total external ventricular power Wt, from 1452(306) to 1121(135) mW; and steady external power Ws, from 1251(310) to 939(119) mW; but did not reduce Zc, fo, Cs, Cm, Cd, and oscillatory external power Wo. CONCLUSIONS: The results indicate that (1) the stiffness of proximal aorta and vascular tone of peripheral arterioles are higher in hypertensives than in normotensives; (2) in hypertensive subjects, sublingual administration of nifedipine reduces the arterial pressure and peripheral arteriolar tone, but not the stiffness of proximal aorta; (3) the decrease in total external ventricular power in hypertensives treated with nifedipine results from a reduction in the steady, but not the oscillatory, component of hydraulic external ventricular power.

Aorta

The influenza resistance murine Mx1 gene is constitutively expressed in the epithelia of the gastrointestinal, respiratory and uterine tracts.

The murine Mx1 gene confers specific resistance against influenza in the inbred A2G mice and in vitro have been shown to be inducible with type I but not type II interferons. Contrary to expectation, we found by in situ hybridisation widespread Mx1 expression along the epithelia of the gastrointestinal, uterine and respiratory tracts in uninduced A2G mice. Several lines of evidence, including further enhancement of Mx1 expression during organ culture and gnotobiotic mice analyses, indicated that this apparent constitutive epithelial Mx1 expression was a locally induced response to stimuli present in the respective lumina. This phenomenon may be a feature found in other interferon-inducible and interferon genes.

Animals