Electronic properties and phase transitions of RbC60 and CsC60: Investigation by NMR spectroscopy.
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Biomedical subjects
Publications and source records attributed to Q Zhu.
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We employed hyperosmotic concentrations of penetrating cryoprotective agents (CPA) to store the isolated rat hearts unfrozen at subzero temperatures. The effect of acute exposure to CPA was assessed by flushing the hearts with CP-14, a cardioplegic solution, containing methanol (MeOH), ethanol (EtOH), ethylene glycol (EG), or propylene glycol (PG) for 2 min and reperfusing immediately with Krebs-Henseleit buffer in a working-heart model. The maximal doses that did not cause irreversible suppression of heart function were: MeOH, 1.78 M; EtOH, 1.27 M; EG, 0.84 M; and PG, 0.87 M. For nonfreezing storage, the hearts were flushed with CP-14 containing the highest tolerable concentrations of MeOH, EtOH, EG, or PG, stored for 6 h at -3.7, -2.8, and -1.4 degrees C, respectively, and then reperfused. Control cardiac output (CO) was 76.2 +/- 1.8 ml/min. Post-reperfusional recovery of CO was 86% in MeOH hearts, 82% in EtOH hearts, 76% in EG hearts, and 79% in PG hearts. Thus MeOH offered not only the least cardiac-suppressing effect but the lowest nonfreezing storage temperature. When storage time was extended, recovery and myocardial ATP level decreased with time in hearts flushed with CP-14 + 1.78 M MeOH and stored at -3.7 degrees C. The decay of function was faster than the decay of ATP level, suggesting energy was better preserved than function. The low return of function, however, may be related to CPA toxicity, osmotic stress, and ischemia/reperfusion injury. Nonfreezing storage at subzero temperatures using these CPAs may provide a novel approach to long-term cardiac preservation.
The basal cell in airway epithelium plays a major role in attachment of ciliated and nonciliated columnar cells to the basal lamina. As the airway grows in diameter and the columnar epithelium in height, the number of basal cells and the amount of tonofilaments (cytokeratin filaments) and anchoring junctions increase. In this way they maintain a constant attachment strength between the increased volume of the epithelium and the basal lamina. The purpose of this study was to determine which cytokeratins (CKs) are expressed in growing basal cells of the rat and demonstrate where they are localized in the cytoskeleton. Sprague Dawley rats 10, 30 and 90 days of age were used in this study. For light microscopy, tracheal samples were fixed in 95% alcohol or 4% formalin for 2 hr and then embedded in paraffin. For electron microscopy, the tracheal samples were placed in 20 mM EDTA in HBSS media minus Ca++ and Mg++ at pH 7.4 for 60 min to permeabilize the cells and expose the intracellular structures. Antibodies to cytokeratins 7, 8, 10, 13 and 18 did not react to basal cells at any age studied. Antibodies to CKs 5 + 8, 14, 16 + 13, and 19 gave a positive reaction with basal cells at each age studied. Immunogold particles representing antibodies to CK 14 were heavily distributed over intermediate filaments making up the cytoskeleton. Both CK 16 + 13 and 19 were also over intermediate filaments but at a much lower density.(ABSTRACT TRUNCATED AT 250 WORDS)
The primary function of basal cells is to attach columnar epithelium to the basal lamina. They may also have limited stem cell potential, but very little is known of other biological functions. Basal cells lie on the basal lamina beneath the ciliated and secretory cells and do not reach the surface of the epithelium. The position of the cell beneath the ciliated and secretory cell epithelium makes their in situ study difficult. In order to further aid in the study of basal cells, we have developed an in situ preparation technique in which ciliated and secretory cells are removed. Treatment of rat tracheas with a 20 mM Na2 EDTA solution, pH 7.4, results in partial removal of columnar epithelium from the basal lamina. The percent of denuded columnar epithelial cells per mm of basal lamina is 43.9 +/- 7.8% at 60 min, 47.6 +/- 8.4% at 90 min, and 52.6 +/- 2.7% at 120 min. The viability of the exposed basal cells was the same at both 60 and 90 min of treatment (79.4 +/- 7.8 and 78.0 +/- 8.5, respectively). Morphologically, the exposed basal cells are attached to the basal lamina by hemidesmosomes and are similar to those in the intact animal.
Amoebae of the eukaryotic microorganism Dictyostelium discoideum were found to contain an interconnected array of tubules and cisternae whose membranes were studded with 15-nm-diameter "pegs." Comparison of the ultrastructure and freeze-fracture behavior of these pegs with similar structures found in other cells and tissues indicated that they were the head domains of vacuolar-type proton pumps. Supporting this identification, the pegs were observed to decorate and clump when broken amoebae were exposed to an antiserum against the B subunit of mammalian vacuolar H(+)-ATPase. The appearance of the peg-rich cisternae in quick-frozen amoebae depended on their osmotic environment: under hyperosmotic conditions, the cisternae were flat with many narrow tubular extensions, while under hypo-osmotic conditions the cisternae ranged from bulbous to spherical. In all cases, however, their contents deep etched like pure water. These properties indicated that the interconnected tubules and cisternae comprise the contractile vacuole system of Dictyostelium. Earlier studies had demonstrated that contractile vacuole membranes in Dictyostelium are extremely rich in calmodulin (Zhu, Q., and M. Clarke, 1992, J. Cell Biol. 118: 347-358). Light microscopic immunofluorescence confirmed that antibodies against the vacuolar proton pump colocalized with anti-calmodulin antibodies on these organelles. Time-lapse video recording of living amoebae imaged by interference-reflection microscopy, or by fluorescence microscopy after staining contractile vacuole membranes with potential-sensitive styryl dyes, revealed the extent and dynamic interrelationship of the cisternal and tubular elements in Dictyostelium's contractile vacuole system. The high density of proton pumps throughout its membranes suggests that the generation of a proton gradient is likely to be an important factor in the mechanism of fluid accumulation by contractile vacuoles.
AD169, a well-characterized laboratory strain of human cytomegalovirus (HCMV), was used to establish an animal model of progressive HCMV chorioretinal disease by injection of 10(5) pfu into the rabbit vitreous. Chorioretinal, vitreous, and pulmonary disease were monitored by HCMV recovery, clinical observation, antigen localization, and histopathology. Vitritis and focal areas of immune cellular infiltrates were seen in inner retinal layers on days 2-4 after inoculation. Disease progressed with more severe vitritis and to involve the outer retinal layers in areas of mixed monocytic cellular infiltrates, retinal destruction, choroidal edema, and congestion. HCMV was recovered from chorioretinal cell sonicate cultures in titers ranging from 10(4) to 10(5) pfu during peak disease, and HCMV antigens were detected focally by immunofluorescence in retinal layers on days 2 and 4 after inoculation. A rabbit model of HCMV chorioretinitis similar to human CMV disease allows investigation of HCMV pathogenesis and new antiviral therapies and evaluation of immune system modulation of the HCMV ocular infection.
This paper describes the development of an adaptive electric current tomography system which contains a novel front-end analog architecture. Programmable voltage sources have been used to deliver currents into the study object and to avoid the difficulties of obtaining high quality current sources. Through inverting an admittance matrix, the system is capable of achieving a desired current drive pattern by applying a computed voltage pattern. The tomograph, operating at 9.6 kHz, comprises 32 driving electrodes and 32 voltage measurement electrodes. The study of system noise performance shows high SNR in the data acquisition which is enhanced by a digital demodulation scheme. In vitro reconstruction images have been obtained with the data collected by the tomograph.
In amoebae of the eukaryotic microorganism Dictyostelium discoideum, calmodulin is greatly enriched on membranes of the contractile vacuole complex, an osmoregulatory organelle. Antibodies specific for Dictyostelium calmodulin were used in the present study to immunolocalize the contractile vacuole complex in relation to the Golgi complex (detected with wheat germ agglutinin) and the microtubule organizing center (MTOC, detected with anti-tubulin antibodies). Cells were examined throughout the cell cycle. Double-staining experiments indicated that the contractile vacuole complex extended to the MTOC in interphase cells, usually, but not always, overlapping the Golgi complex. In metaphase and anaphase cells, the Golgi staining became diffuse, suggesting dispersal of Golgi membranes. In the same mitotic cells, anti-calmodulin antibodies labeled numerous small cortical vacuoles, indicating that the contractile vacuole complex had also become dispersed. When living mitotic cells were examined, the small cortical vacuoles were seen to be active, implying that all parts of the Dictyostelium contractile vacuole complex possess the ability to accumulate fluid and fuse with the plasma membrane. In contrast to observations reported for other types of cells, anti-calmodulin antibodies did not label the mitotic spindle in Dictyostelium. Despite this difference in localization, it is possible that vacuole-associated calmodulin in Dictyostelium cells and spindle-associated calmodulin in larger eukaryotic cells might perform a similar function, namely, regulating calcium levels.
As a kind of anti-inflammatory protein fractionated and purified from the larva of parasa sinica, CCP (ip) has a significant anti-inflammatory effect on ear odema induced by croton oil in mice. Its ID50 is 1.6mg/kg, but a dose of 2.5mg/kg can also significantly inhibit the rat ankle odema induced by carrageenan and egg white.
The authors report the results of a parallel study between the three kinds of evoked potentials (EP) i.e. visual, brainstem auditory and somatosensory evoked potentials (VEP, BAEP and SEP) and psychometric tests in 30 posthepatitic cirrhotic patients. The results were compared with those from 20 peptic ulcer patients in healing stage. Psychometric tests were abnormal in 65.5% of the cirrhotic and in none of the peptic ulcer patients. The three kinds of evoked potentials were abnormal in 58.6% of the cirrhotic patients and in 5% of the peptic ulcer patients. The VEP, BAEP and SEP was abnormal in 13.3%, 10% and 50% of the cirrhotics respectively. The results of this study showed: of the three kinds of evoked potentials, SEP is the most sensitive one. The evoked potentials and the performance psychometric tests are complementary in the diagnosis of subclinical hepatic encephalopathy.
Fifty patients with thyroid nodules were examined by high-resolution real-time ultrasonography (US) before surgery. Of this series, the sonographic features of 54 nodules in 41 cases were classified according to internal echogenicity, cystic components, margin, sound transmission, anterior extra-thyroid extension, and peripheral "halo" sign. Except for the last, the differences between benign and malignant nodules were statistically significant (P < 0.05, P < 0.01 or P < 0.001). The comparisons of US and palpation examination (PE) with postoperative pathological findings were undertaken in these 50 cases, respectively. The sensitivity and accuracy for US were much higher than those for PE (sensitivity, 97% vs. 77%; accuracy, 97% vs. 76%). Our results show that by analyzing some sonographic characteristics, it may be possible to differentiate benign from malignant lesions and owing to its higher sensitivity and accuracy, ultrasonography is a better choice of the scrutiny into thyroidal gland.
To analyze the importance of cell surface-associated molecules in modulating the immune response by facilitating T/B cell interaction, we used the T cell-dependent antigen, bacteriophage phi X174. Taking advantage of "experiments of nature", we studied specific antibody synthesis in patients with deficiencies of complement components or of the adhesion molecule CD11/CD18 (leukocyte adhesion defect, LAD) and guinea pigs and dogs with early complement component deficiency. Following intravenous injection of bacteriophage phi X174 into normal subjects or animals, a primary response consisting of IgM, a secondary response consisting of IgM and IgG, and a tertiary, predominantly IgG response can be distinguished. Patients and guinea pigs deficient of early complement component and LAD patients responded to repeated phage immunization with depressed antibody titers, lack of or inadequate amplification, and failure to switch from IgM to IgG, suggesting a defect in generating antigen-specific memory cells. Several mechanisms have to be considered: (i) The complement portion of the antigen-antibody complement complex facilitates the accumulation and trapping of antigen in lymphoid organs, thus improving the response to Ag at low concentrations. (ii) Immune complexes preferentially bind to antigen-specific B cells, cells expressing Fc receptors, or CR2 and CR3, the receptors for C3bi. (iii) The weak binding established between the MHC-II/Ag complex and the TCR complex is strengthened through the binding of several adhesion molecule pairs. (iv) Receptor-ligand binding initiates activation signals. The concept of binding/signaling via interacting molecules is further supported by the observation that mAb 60.3, recognizing the beta chain of CD11/CD18, blocks in vitro synthesis of antibody to bacteriophage by primed PBMC.
This study investigated the effect of thawing rate on the preservation of frozen isolated rat hearts. The hearts were flushed with a hyperosmotic cardioplegic solution, CP-14/EtOH (1.15 Osm/kg), frozen at a rate of 0.18 degree C/hr for 6 h to -3.2 degrees C. Thereafter, the hearts were thawed at rates ranging from 0.08 to 1.1 degrees C/min for 1 to 14 min until the heart temperature reached -2.1 degrees C, the melting point (MP) of the flush solution; then they were held at -1 degree C for 11 to 24 min so that the total thaw time was 25 min. Post-thaw function was assessed by working reperfusion and expressed as percentage of unstored control function. Cardiac output (CO) and other hemodynamic performance showed biphasic responses to the thaw rate. At 0.08 degree C/min rate, CO recovered to 29.1 +/- 4.1 ml/min (40.8 +/- 5.8% of control). Thawing at 0.13 degree C/min enhanced the recovery of CO to 60.5 +/- 4.9%. Between 0.13 and 0.34 degree C/min, recovery was statistically insignificant. Faster thawing at 0.59 and 1.1 degrees C/min caused progressively less recovery. Overall, 0.13 degree C/min offered the highest recovery. In conclusion, function in slowly frozen heart is intimately affected by the thawing rate; there was an optimal intermediate thawing rate and both too slow and too fast thawing were detrimental.
mAbs specific for calmodulin were used to examine the distribution of calmodulin in vegetative Dictyostelium cells. Indirect immunofluorescence indicated that calmodulin was greatly enriched at the periphery of phase lucent vacuoles. The presence of these vacuoles in newly germinated (non-feeding) as well as growing cells, and the response of the vacuoles to changes in the osmotic environment, identified them as contractile vacuoles, osmoregulatory organelles. No evidence was found for an association of calmodulin with endosomes or lysosomes, nor was calmodulin enriched along cytoskeletal filaments. When membranes from Dictyostelium cells were fractionated on equilibrium sucrose density gradients, calmodulin cofractionated with alkaline phosphatase, a cytochemical marker for contractile vacuole membranes, at a density of 1.156 g/ml. Several high molecular weight calmodulin-binding proteins were enriched in the same region of the gradient. One of the calmodulin-binding polypeptides (molecular mass approximately 150 kD) cross-reacted with an antiserum specific for Acanthamoeba myosin IC. By indirect immunofluorescence, this protein was also enriched on contractile vacuole membranes. These results suggest that a calmodulin-binding unconventional myosin is associated with contractile vacuoles in Dictyostelium; similar proteins in yeast and mammalian cells have been implicated in vesicle movement.
The effects of coal tar pitch (CTP) on the tracheobronchial mucosa of Wistar rats were studied. Three groups of animals received 10 weekly intratracheal instillations of CTP at the cumulative doses of 6.48, 136.56 and 200 mg respectively. The control group of rats received 10 weekly intratracheal instillations of charcoal powder at a cumulative dose of 20 mg. The study in which the animals were killed serially revealed that CTP had conspicuous damage on the respiratory system of rats, especially on the bronchiolo-alveolar areas. The lesions induced by CTP ranged from hyperplastic, metaplastic and dysplastic changes to extensive cancers. These lesions were usually multifocal, and were more severe in the rats receiving higher dosages of CTP. The deposition of CTP particles within or adjacent to these lesions could be readily identified. Lung cancers occurred in 12.5% (4/32) and 25% (10/40) of the rats treated with 136.56 and 200 mg of CTP, whereas no tumors were found in control rats and the rats that received 6.48 mg of CTP. The overall cancer incidence significantly related to the cumulative dose of CTP. The histological types of lung cancers consisted of squamous cell carcinomas (10 out of the 14 lung cancers), adenocarcinoma (1/14), and combined squamous and adenocarcinomas (3/14). The development of CTP-induced rat lung cancers appears to derive from the hyperplasias of bronchiolo-alveolar epithelium, and processing stages of squamous metaplasias and/or dysplasias to carcinomas. The present results confirmed the carcinogenic effects of CTP on the respiratory system of rats, and provided experimental evidence for human lung carcinogenesis, particularly in those occupationally exposed to coal tars or tar products.
Adenosine deaminase was infused into isolated perfused guinea pig hearts to determine its effect on myocardial adenosine levels. The enzyme was administered during constant coronary flow perfusion at 6.11 +/- 0.36 ml.min-1.g-1. Venous adenosine was measured in samples of pulmonary artery effluent; epicardial and endocardial adenosine were measured with the porous nylon disk technique. Infusion of adenosine deaminase at 2.4 and 4.8 U/ml produced adenosine deaminase activity of 0.92 +/- 0.09 and 2.33 +/- 0.15 U/ml, respectively, in epicardial fluid and 1.93 +/- 0.28 and 4.84 +/- 0.47 U/ml, respectively, in endocardial fluid. Aortic pressure was unchanged by infusion of adenosine deaminase at both infusion rates. Adenosine deaminase (data from both infusion rates pooled) reduced epicardial adenosine from 0.327 +/- 0.028 to 0.139 +/- 0.022 microM, endocardial adenosine from 4.61 +/- 0.42 to 1.64 +/- 0.20 microM, and venous adenosine from 0.017 +/- 0.02 to 0.003 +/- 0.001 microM. The data indicate that infused adenosine deaminase reaches the epicardial and endocardial interstitial fluid (ISF) compartments. The absence of any effect on coronary pressure suggests that adenosine may not be involved in resting basal coronary tone. The presence of significant residual adenosine despite adenosine deaminase infusion indicates that adenosine production in the unstressed isolated guinea pig heart exceeds the degradative capacity of infused adenosine deaminase. Previous studies in which it was assumed that almost all of the endogenous adenosine is inactivated by the infusion of adenosine deaminase should be reevaluated in light of these observations.