Studies of topological distributions of inclusive three- and four-jet events in p-barp collisions at s=1800 GeV with the D0 detector.
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Biomedical subjects
Publications and source records attributed to L Chen.
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BACKGROUND: A viral etiology has been suspected in papillomatosis of the external auditory canal (PEAC), but virus particles have not been detected so far, although they are easily demonstrable in skin warts. The purpose of the study was to solve this discrepancy by the use of polymerase chain reaction (PCR). MATERIALS AND METHODS: Specimens from the external auditory canal of 14 patients with PEAC, but no human papilloma virus infection of the genital areas, were examined histologically by light and electron microscopy, as well as by PCR to detect viral DNA. RESULTS: Histologically, papillomatosis was present in all specimens. Vacuolated cells were found in the upper part of the stratum malpighii in five cases. On electron microscopy, the numbers of perichromatin and interchromatin granules were increased, but no viral granules were observed. In all specimens, DNA of HPV 6 was detected using PCR, but there was no evidence of DNA of other HPV. CONCLUSIONS: Papilloma of the external auditory canal is produced by infection with HPV 6.
Chlorophyll (Chl) and bacteriochlorophyll (Bchl) have been made water soluble by transesterification with serine (Ser) at the propionyl residue and tested as potential reagents for photodynamic therapy (PDT). Photocytotoxicity of the conjugates Chl-Ser and Bchl-Ser in M2R mouse melanoma was tested in cell cultures. Tissue uptake and clearance of the photosensitizers in CD1 nude and C57B1 mice implanted with M2R tumors are described. Photocytotoxicity in cell cultures was determined microscopically and by [3H]thymidine incorporation. The LD50 values in vitro were 0.05-0.1 microM for both sensitizers while that of the commercially available hematoporphyrin derivative (HPD, Photosan) was over 100 times higher for the same light intensity (45 mW/cm2). Pigment concentrations were determined fluorometrically in acetone extracts of the tissues of interest at different times after intraperitoneal injection of 20 mg pigment/kg body weight. The distribution pattern of Chl-Ser in the different tissues resembled that reported for Photofrin, chlorin and bacteriochlorin derivatives. Clearance from normal tissues was essentially completed within 16 h for Bchl-Ser and 72 h for Chl-Ser with mean half-lives (t 1/2) of about 2 and 7 h, respectively. In contrast, the clearance rates of these pigments and their metabolites from melanoma tumor tissue were significantly longer: t 1/2 = 20 h for Chl-Ser and 15 h for Bchl-Ser and metabolites. The clearance rates showed biphasic or single exponential decay patterns in normal tissues and in tumors, respectively. Cumulatively the high phototoxicity, simple mode of delivery and fast tissue clearance rates reported here suggest that polar conjugates of Chl and Bchl promise to be highly effective PDT reagents.
The purpose of the present study was to provide a quantitative description of two-tone rate suppression boundaries in normal chickens. The boundaries were measured in 249 cochlear ganglion neurons using a tone 20 dB above threshold at the characteristic frequency (CF). The boundaries were present in 90.4% of neurons either on both sides or only one side of CF but more frequently above CF than below CF. The best suppression thresholds were positively correlated with and, on the average, 19-25 dB higher than CF thresholds. The boundary was farther from CF and shallower below CF than above CF. The boundary slope varied slightly with CF threshold and the tuning curve slope. These results are generally consistent with previous reports from mammals except that: (1) the boundary below CF did not follow and lie above the tuning curve flank; (2) the average best suppression threshold was slightly lower below CF than above CF; (3) the boundaries below and above CF were not particularly asymmetrical.
In Bacillus subtilis, hydrogen peroxide induces the synthesis of catalase (KatA), alkyl hydroperoxide reductase (AhpCF), and a DNA-binding protein of the Dps family (MrgA). KatA, AhpCF, heme biosynthesis enzymes, and MrgA are also induced upon entry into stationary phase under conditions of iron and manganese limitation. In an effort to define the peroxide regulon repressor, PerR, we used mini-Tn10 mutagenesis to identify loci affecting the regulation of mrgA. From this screen, we isolated two mini-Tn10 insertions in ahpC, the gene encoding the small subunit of AhpCF, that increase the transcription of mrgA-lacZ even in iron-supplemented minimal medium. Indeed, these ahpC::Tn10 insertions lead to elevated expression from all peroxide regulon promoters, including those for mrgA, katA, hemAXCDBL, and ahpCF. As a result, the ahpC::Tn10 mutants display an increased resistance to H2O2. The ahpCF promoter region contains three sequences similar to the peroxide regulon consensus operator (per box). We demonstrate that the ability of ahpC::Tn10 mutations to derepress mrgA requires aerobic growth. In contrast, a second distinct trans-acting regulatory mutation bypasses this requirement for aerobic growth. Since the peroxide regulon is activated in the absence of AhpCF, which degrades alkyl hydroperoxides, we propose that organic hydroperoxides may be physiologically relevant inducers in vivo.
Regulated secretion requires the developmental coupling of neuronal or hormonal stimuli to an exocytotic response, a multistep pathway whose appearance may be linked with cellular adhesion to the newly formed exocrine cell basement membrane. We screened for adhesion-associated coupling activity using lacrimal acinar cells and have identified "BM180", a novel basement membrane protein enriched in guanidine HCl extracts of lacrimal and parotid exocrine secretory glands. BM180 resides primarily in a previously inexamined lower molecular-mass basement membrane peak (peak 2) that contains cell adhesion activity inhibitable with the anti-BM180 monoclonal antibody 3E12. Removal of peak 2 by gel filtration or preincubation of basement membrane with 3E12 decreased regulated peroxidase secretion by one-half without affecting constitutive secretion or the amount of cellular peroxidase available for release. Adding back peak 2 restored regulated secretion in a dose-dependent and 3E12-inhibitable manner and suggested a synergistic relationship between BM180 and laminin 1. BM180 has a mobility of 180 and 60 kDa in the absence or presence of dithiothreitol, respectively, and shows no immunological identity by competitive enzyme-linked immunosorbent assay with laminin 1, collagen IV, entactin, fibronectin, BM-40, perlecan, or vitronectin. We propose that BM180 is an important resident of certain glandular basement membranes where it interacts with the cell surface, thereby possibly signaling the appearance of a transducing element in the stimulus-secretion coupling pathway.
We have previously shown that whole-cell L-type Ca2+ current that was stimulated through beta-adrenergic receptors was negatively modulated by alpha 1-adrenergic activation. In the present study, we investigated the kinetic basis of this modulation at the single-channel level in adult rat ventricular myocytes using Ba2+ as the charge carrier. Unitary current sweeps were evoked by 300-ms depolarizing pulses to 0 mV, from a holding potential of -50 mV at 0.5 Hz. During control conditions, the ensemble-averaged current amplitude was 0.18 +/- 0.01 pA (n = 7). To achieve beta-adrenergic stimulation (beta effect), cells were superfused with norepinephrine (10 mumol/L) in the presence of prazosin (10 mumol/L), an alpha 1-adrenergic blocker. beta-adrenergic stimulation enhanced ensemble-averaged current (from 0.18 +/- 0.01 to 0.75 +/- 0.04 pA, P < .05, n = 7), increased the open-time constants, and decreased the closed-time constants. To activate alpha 1-receptors while maintaining the beta-adrenergic stimulation, cells were superfused with norepinephrine alone (alpha 1 + beta effects). alpha 1-adrenergic activation reduced ensemble-averaged current (from 0.75 +/- 0.04 to 0.41 +/- 0.03 pA, P < .05, n = 7), decreased open-time constants, and increased closed-time constants. alpha 1-adrenergic activation also inhibited ensemble-averaged currents stimulated by a low concentration (10 mumol/L) of 8-bromo-cAMP but not by (-)Bay K 8644 (1 mumol/L). Calphostin C (1 mumol/L), a specific inhibitor of protein kinase C, attenuated alpha 1-adrenergic inhibition on beta-adrenergic-stimulated unitary currents. We conclude that alpha 1-adrenergic activation exerts an inhibitory effect on beta-adrenergic-stimulated unitary Ba2+ current at the single-channel level. The shortening of the open-time and the lengthening of the closed-time constants and the increase in blank sweeps may explain the inhibition of the Ca(2+)-channel activity and the reduction in whole-cell Ca2+ current previously reported. This inhibition is in part mediated through the protein kinase C pathway.
In order to test the hypothesis that ozone (O3)-induced changes in lung function and respiratory tract injury/inflammation are greater in subjects with asthma than in normal subjects, we exposed 18 asthmatic subjects, on separate days, to O3 (0.2 ppm) and filtered air for 4 h during exercise. Symptom questionnaires were administered before and after exposure, and pulmonary function tests (FEV1, FVC, and specific airway resistance [SRaw]) were performed before, during, and immediately after each exposure. Fiberoptic bronchoscopy, with proximal airway lavage (PAL) of the isolated left main bronchus and bronchoalveolar lavage (BAL; bronchial fraction, the first 10 ml of fluid recovered) of the right middle lobe, was performed 18 h after each exposure. The PAL, bronchial fraction, and BAL fluids were analyzed for the following endpoints: total and differential cell counts; total protein, lactate dehydrogenase (LDH), fibronectin, interleukin-8 (IL-8), granulocyte-macrophage colony-stimulating factor (GM-CSF), myeloperoxidase (MPO), and transforming growth factor-beta (TGF beta 2) concentrations. We found a significant O3 effect on FEV1, FVC, SRaw (p < 0.04) and lower respiratory symptoms (p < 0.001) for the asthmatic subjects. Ozone exposure also significantly increased the percent neutrophils in PAL (p < 0.01); percent neutrophils, total protein, and IL-8 in the bronchial fraction (p < 0.001, p < 0.05, and p < 0.01, respectively); and the percent neutrophils, total protein, LDH, fibronectin, IL-8, GM-CSF, and MPO in BAL (p < 0.001, p < 0.01, p < 0.01, p < 0.001, p < 0.05, p < 0.01, and p < 0.001, respectively) for the asthmatic subjects. There were no significant differences in the lung function responses of the asthmatic subjects in comparison with a group of normal subjects (n = 81) previously studied using an identical protocol, although there was a trend toward a greater O3-induced increase in SRaw in the asthmatic subjects (p < 0.13). In contrast, the asthmatic subjects showed significantly greater (p < 0.05) O3-induced increases in several inflammatory endpoints (percent neutrophils and total protein concentration) in BAL as compared with normal subjects who underwent bronchoscopy (n = 20). Our results indicate that asthmatic persons may be at risk of developing more severe O3-induced respiratory tract injury/inflammation than normal persons, and may help explain the increased asthma morbidity associated with O3 pollution episodes observed in epidemiologic studies.
OBJECTIVE: This study investigated the role of maternal smoking during pregnancy in the etiology of attention deficit hyperactivity disorder (ADHD). METHOD: Subjects were 6-17-year-old boys with DSM-III-R ADHD (N = 140) and normal comparison subjects (N = 120) and their first-degree biological relatives. Information on maternal smoking was obtained from mothers in a standardized manner by raters who were blind to the proband's clinical status. RESULTS: Twenty-two percent of the ADHD children had a maternal history of smoking during pregnancy, compared with 8% of the normal subjects. This positive association remained significant after adjustment for socioeconomic status, parental IQ, and parental ADHD status. Significant differences in IQ were found between those children whose mothers smoked during pregnancy and those whose mothers did not smoke (mean IQ = 104.9, SD = 12.3, and mean = 115.4, SD = 12.2, respectively). CONCLUSIONS: These findings suggest that maternal smoking during pregnancy is a risk factor for ADHD. If confirmed, these findings will stress the importance of programs aimed at smoking prevention in nonsmoking women and smoking cessation in smoking women of childbearing age.
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BACKGROUND: To study the transient outward current (I(to)) investigators often use sodium-free external solution to minimize the possible contamination of I(to) by sodium current. Removal of extracellular sodium creates reversal of sodium gradient and thus possibly contributing to I(to) mainly at positive potentials. METHODS AND RESULTS: To address this issue, whole-cell I(to) was recorded in sodium-free choline chloride and cobalt solutions, from rat ventricular myocytes known to exhibit a prominent I(to). Depolarizing pulse to 40 mV from -100 mV holding potential every 10 seconds elicited a fast activating and time-dependent inactivating components. The activation of I(to) was fast and complete within 10 ms at 40 mV, and the decay was rapid over the first 100 ms of the pulse and slower thereafter. External superfusion of the cell with 50 µM tetrodotoxin reversibly reduced I(to) amplitude by 25% from 1.47 +/- 0.2 to 1.1 +/- 0.3 nA (P <.04, n = 9). When sea anemone toxin (ATXII), known to selectively enhanced I(Na) by causing a delay in the inactivation gate, is applied to the cell, I(to) amplitude increased in a time- and dose-dependent manner (EC(50) =.86.4 nM). ATXII (100 nM) dramatically increased I(to) amplitude at all voltages between -20 and 60 mV (from 1.51 +/- 0.4 to 3.35 +/- 0.8 nA at 40 mV, P <.003, n = 12). Superfusion of cells with 5 mM 4-AP resulted in 82% reduction in I(to) amplitude at 40 mV (from 1.95 +/- 0.5 to 0.37 +/- 0.2 nA, P <.02, n = 8). Addition of ATXII to 4-AP containing solution increased peak I(to) by 965% (from 0.37+/-0.2 to 3.95 +/- 0.9, n = 8, P <.0003). However, in 11 other cells, addition of tetrodotoxin (50 µM) to the ATXII-containing solution blocked ATXII-induced outward current (from 3.51 +/- 0.64 nA to 1.60 +/- 0.17 nA, P <.05). The conductance (G(Ito)) was calculated by dividing peak I(to) by (Vm-E(K)), with an E(K) of -75 mV. G(Ito) was increased at all voltages (greater than -40 mV). Normalized G(Ito) was fitted by Boltzmann equation and ATXII did not significantly modify V(0.5) and k (from -20.5 +/- 3.9 to -17.0 +/- 3.5 mV for V(0.5), and 12.2 +/- 2.6 to 13.4 +/- 2.1 mV for k, n = 4). Also, atropine (1 µM) did not have any significant effect on I(to) (from 1.92 +/- 0.15 nA to 1.85 +/- 0.25 nA, n = 5). CONCLUSIONS: The results showed that, in sodium-free external solution I(to) is tetrodotoxin but not atropine sensitive. ATXII-induced I(to) increase is 4-aminopyridine insensitive but tetrodotoxin sensitive. These data suggest that outward Na current due to reversal of Na gradient in the absence of external Na contributes to I(to). Caution must be taken when studying kinetics and pharmacology of I(to) in external sodium-free solutions.
Hepatic metastases of malignant tumors is a major problem in the treatment of cancers for which the liver is the most common site for recurrences. In the present study we describe a selective delivery system to the liver which may facilitate specific hepatic targeting of anti-cancer agents. Avidin and streptavidin are two biotin-binding proteins with extreme resistance to proteolytic activity. Trinitrophenyl (TNP) modification of these two proteins resulted in specific accumulation in mouse liver with levels of 40-50 percent per gram tissue (%/g) during a period of several days. The two modified proteins could target to the liver high doses of covalently bound radionuclide iodine-125, a biotinylated ligand such as biotinyl-tyrosine (BT) or large biotinylated carriers such as carboxymethyl dextran (CMdex, 40kDa). Appropriately derivatized dextrans serve as carriers for various chemotherapeutic drugs, as demonstrated here for cis-dichlorodiammineplatinum (CDDP). Specific liver targeting of CDDP complexed to CMdex-TNP-streptavidin could be monitored by flame atomic absorption spectrometry of the Pt metal: High levels of the Pt drug were concentrated in the liver for at least 15hr following its targeted delivery as compared to essentially undetectable levels after administration of the free drug.
The expression of connexin43, the primary gap-junction constituent of glial cells, was evaluated at the messenger RNA and protein levels in different grades of astrocytoma to investigate the relevance of gap junctions in herpes simplex virus-thymidine kinase (HSV-tk)-mediated gene therapy of brain tumors. Transduction of the retroviral-mediated HSV-tk gene into tumor cells with subsequent administration of ganciclovir has recently been used as an experimental therapeutic strategy for treatment of brain tumors. One aspect of this approach is the bystander effect, which augments the efficacy of this therapeutic approach. Glioblastoma cells with minimum levels of connexin43 protein were transfected with a connexin43 complementary DNA. These cells manifested a marked increase in the in vitro bystander effect, supporting the contention that the in vitro bystander effect is a consequence of metabolic cooperation between cells mediated by gap junctions. To assess relative levels of gap-junction protein expression in the relevant tumor type, we examined primary astrocytomas, primary astrocytoma cell cultures, and glioblastoma cell lines. Although most astrocytoma tumor samples expressed connexin43, they differed in the level of expression, with the greatest variation exhibited in high-grade astrocytomas. Primary glioblastoma cell cultures and established glioblastoma cell lines also displayed some variability in connexin43 levels. In aggregate, our results anticipate that glioblastomas will have a varied bystander effect during HSV-tk gene therapy depending on the level of connexin43 expression.
The case of an interdural arachnoid cyst of traumatic origin at the C3-5 level in an 18-year-old man was admitted to our hospital with a 1-year history of progressive weakness in left limbs and numbness below the clavicles is reported. He had had a C-2 fracture at the age of 9 years without definite neurological deficits. Magnetic resonance imaging revealed a cystic lesion in the C3-5 level. Laminectomy was performed, and an interdural cyst was found. Histological examination revealed fibrous thickening of the arachnoid membrane. A cyst located in the interdural space of upper cervical spine is extremely rare.
This paper reports the pathologic and functional changes of the sciatic nerve indirectly injured by firearms and its effect on wound healing. Twelve dogs were used. Their hind legs were shot with 1.03-gram steel spheres, avoiding direct injury to the sciatic nerve. The experimental animals were divided into two groups according to the impact velocity, that is, a 1,500 m/sec high velocity projectile (HVP) group and a 400 m/sec, low velocity projectile (LVP) group as a control. After wounding, the same procedure was followed in all dogs. At intervals, the tissues of the wound track were excised to determine the level of cyclic nucleotides (cAMP, cGMP), secondary messengers of neurotransmitters, the activity of acetylcholinesterase (AChE), and the content of hydroxyproline, which indicates the degree of wound healing. The sciatic nerve next to the wound track was excised for light and electron microscopic study. The neurofilament in the sciatic nerve was examined by immunohistochemistry with monoclonal neurofilament-200 antibody. The results showed that the indirect injuries to the sciatic nerves of the HVP group were more severe than in those in the LVP group and included naked axons, stripped myelin sheaths, widened nodes of Ranvier, and decrease in the numbers of neurofilaments. At 6 hours, cAMP content in the HVP group was significantly lower than that in the LVP group. It began to rise and approached to the level of LVP group at 10 days. The cGMP content, activity of AChE, and hydroxyproline content of the HVP group were significantly lower than those of the LVP group. The results suggested that the indirect injury to sciatic nerves, especially to the cytoskeleton, after HVP wounds may induce the disturbance of an axoplasmic flow and influence the metabolism of trophic factors such as cAMP, cGMP, and AChE. All changes could contribute to the delay in wound healing.
Coronary artery often reoccludes after therapy of acute myocardial infarction with recombinant tissue plasminogen activator rt-PA, most likely due to in situ thrombin generation during thrombolysis. Previous studies with high molecular weight thrombin inhibitors, such as hirudin, have shown variable improvement in the frequency of sustained thrombolysis. This study was conducted to examine the modulation of thrombolysis, indices of thrombin generation and activated partial thromboplastin time (APTT) by a novel low molecular weight direct thrombin inhibitor, inogatran. A stable occlusive intracoronary thrombus was created in 19 dogs. Nine dogs were given an intravenous bolus of saline (group A), and 5 dogs were given inogatran (group B) followed by rapid infusion of rt-PA (1 mg/kg over 20 min), whereas saline or inogatran was continuously infused for 2 hr. Five other dogs were given inogatran (bolus and continuous infusion) only after thrombolysis by rt-PA was obtained (group C). Time to reflow was similar in all dogs. None of the reperfused coronary arteries reoccluded in group B dogs (vs. 75% and 40% reocclusion rates in groups A and C, respectively, P < .02). Accordingly, the mean duration of reflow was > 120 min in group B dogs (vs. 39 +/- 7 and 44 +/- 14 min in group A and C dogs, respectively, P < .05). After infusion of inogatran, APTT was increased to 1.6 to 1.9 times the base-line value, and the changes in APTT were similar in group B and C dogs. Thrombin generation and activity, assessed by thrombin-antithrombin complex and fibrinopeptide A levels, increased in all dogs during thrombus formation. The increase in thrombin-antithrombin complex and fibrinopeptide A levels during thrombolysis was not evident in group B dogs. These data show that direct thrombin inhibition with inogatran, when initiated before rt-PA, results in sustained thrombolysis and only a modest increase in APTT. However, inogatran given after thrombolysis only partially prevents reocclusion because large amounts of thrombin generation occur during the early stages of thrombolysis.
Previous reports indicated that treatment of rats with green tea or black tea extracts increased CYP1A2 activity, but such an induction was not observed with decaffeinated green tea in our preliminary study. Herein we report a comparative study on the induction of CYP1A2 with different tea preparations and caffeine as an inducer. When green tea (2%) or black tea (2%) was given to male Fischer 344 rats as the sole source of drinking fluid for 21 days, a 2.4- or 2.7-fold induction, respectively, of CYP1A2-dependent O-methoxyresorufin demethylase (MROD) activity in liver microsomes was observed. Treating rats with caffeine (0.04%) also resulted in an 1.9-fold increase in the MROD activity, but decaffeinated green tea (0.8%) did not cause such an induction. Rats treated with green tea (2%) or caffeine (0.055%) as the sole source of drinking fluid for 1, 3, and 7 days also showed comparable induction (from 1.7- to 2.1-fold) of the MROD activity. The induction was also shown by intragastric administration of caffeine (100 mg/kg). The induced MROD activity caused by consumption of green tea, black tea, and caffeine corresponded to the increase in liver microsomal CYP1A2 protein, as determined by immunoblot analysis. The concentrations of tea polyphenols and caffeine in plasma were also measured. Close correlation of the increase in the MROD activity was observed only with the plasma caffeine level (r = 0.736, n = 10, p = 0.015), not with the combined tea polyphenol level (r = 0.058, n = 6, p = 0.913). The present study establishes caffeine as an inducer of CYP1A2 and demonstrates that caffeine, not tea polyphenols, is the component in tea responsible for the induction of this enzyme.
Surgical samples from 40 cases of primary lung cancer were studied by in situ hybridization and polymerase chain reaction (PCR) to detect HPV-DNA in different types of lung cancer. In addition, PCR-RFLP was used to examine mutation of p53 exon 7. The results showed that HPV-DNA positive rate in lung cancer was 55% (22/40 cases), including SCLC (9/9 cases), squamous cell carcinomas (8/16 cases), and adenocarcinomas (5/12 cases). Amplification of p53 exon 7 was seen in 5 of 22 HPV-DNA positive cases. RFLP analysis showed that p53 exon 7 mutation was present in two cases with gene amplification. In HPV-DNA negatives there was only one case with exon 7 amplification but no mutation. SCLC and squamous carcinoma had higher HPV infection rate than other types of lung cancer. The results suggest some relation between HPV infection and p53 gene mutation.