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Biomedical subjects

J Hua

Publications and source records attributed to J Hua.

At least 109 records · Page 6Linked to original sources
PubMed1993

Anti-bombesin monoclonal antibodies modulate fetal mouse lung growth and maturation in utero and in organ cultures.

Fetal pulmonary neuroendocrine cells (PNECs) contain abundant gastrin-releasing peptide (GRP, mammalian bombesin-like peptide [BLP]). Previously, addition of bombesin resulted in increased fetal lung growth and maturation in utero and in organ cultures. A monoclonal antibody (mAb) to bombesin (2A11) blocked baseline automaturation of lung organ cultures in serum-free medium. In the present study, we analyze lung development following daily in utero administration of 2A11 from gestational days 15-18. Fetal lung treated with 2A11 and then harvested on day 18 demonstrated a dose-dependent decrease in surfactant phospholipid synthesis compared to controls treated with MOPC, an unreactive mAb. However, 2A11-treated fetal lung harvested on day 17 showed paradoxical increases in 3H-choline incorporation into saturated phosphatidylcholine, 3H-thymidine incorporation into DNA, and relative numbers of differentiated type II pneumocytes. In serum-containing day 17 lung organ cultures, 2A11 stimulated choline and thymidine incorporation. Since epidermal growth factor (EGF) is the only agent besides bombesin known to stimulate both fetal lung growth and maturation, we added EGF to serum-free cultures and reconstituted the stimulatory effects. A murine EGF receptor mAb (ERA) blocked 2A11-induced lung growth and maturation in serum-containing cultures, and this effect was overcome by adding EGF. In vivo, ERA also blocked stimulatory effects of 2A11 in fetal lung on day 17. These observations suggest that EGF receptor up-regulation may maintain lung growth and maturation if BLP levels are diminished on day 17. Nonetheless, BLPs appear to be involved in lung maturation on day 18, supporting a role for PNECs in normal lung development.

Animals↗
PubMed1993

Comparative analysis of the rotavirus NS53 gene: conservation of basic and cysteine-rich regions in the protein and possible stem-loop structures in the RNA.

NS53, the product of rotavirus gene 5, is an RNA-binding protein that contains a cysteine-rich region and is a component of early replication intermediates. To gain information about the structure of NS53 and its RNA, we determined the nucleotide sequence of gene 5 for the human viruses Wa (serotype 1) and DS1 (2) and the simian virus SA11 (3) (Patton strain) and compared them and their deduced amino acid sequences to those reported for the bovine viruses UK (6) and RF (6), SA11 (3) (Both strain), the human virus Rohivg803, and the group C porcine virus PRV. The results showed that gene 5 for human, simian, and bovine strains have lengths of 1564-1567, 1611, and 1579-1581 nucleotides (nt) and encode proteins of 486, 495, and 491 amino acids, respectively. Comparison of the protein sequences for NS53 among different serotypes showed that they are extremely divergent with many sharing amino acid homologies of only 36-38%. Even NS53 from viruses isolated from the same species possessed relatively poor homology, e.g., DS1 versus Wa was 68%. The first 150 amino acids of NS53 exhibited a greater degree of conservation than the rest of the protein. Near the amino terminus, NS53 contains three basic regions and a cysteine-rich domain, suggesting that this area is responsible for the RNA-binding activity of the protein. Present in the cysteine-rich domain of all group A and C viruses was the motif C-X2-C-X8-C-X2-C-X3-H-X-C-X2-C-X5-C. Although this motif may form one or two zinc fingers, the fact that it is highly conserved indicates that it plays a critical role in the function of protein. Comparison of the nucleotide sequences for gene 5 showed that the entire 5'-noncoding region and the first 24 nt of the NS53 ORF are conserved. RNA-folding predictions suggest that this region of the NS53 mRNA can interact with itself, producing a stem-loop structure similar to that found near the 5'-terminus of the NS35 mRNA. Thus, such structures may be common to all rotavirus mRNAs, perhaps functioning as signals for packaging of RNAs into replication intermediates or regulating mRNA translation.

Amino Acid Sequence↗
PubMed1993

Location of intrachain disulfide bonds in the VP5* and VP8* trypsin cleavage fragments of the rhesus rotavirus spike protein VP4.

Because the rotavirus spike protein VP4 contains conserved Cys residues at positions 216, 318, 380, and 774 and, for many animal rotaviruses, also at position 203, we sought to determine whether disulfide bonds were structural elements of VP4. Electrophoretic analysis of untreated and trypsin-treated rhesus rotavirus (RRV) and simain rotavirus SA11 in the presence and absence of the reducing agent dithioerythritol revealed that VP4 and its cleavage fragments VP5* and VP8* possessed intrachain disulfide bonds. Given that the VP8* fragments of RRV and SA11 contain only two Cys residues, those at positions 203 and 216, these data indicated that these two residues were covalently linked. Electrophoretic examination of truncated species of VP4 and VP4 containing Cys-->Ser mutations synthesized in reticulocyte lysates provided additional evidence that Cys-203 and Cys-216 in VP8* of RRV were linked by a disulfide bridge. VP5* expressed in vitro was able to form a disulfide bond analogous to that in the VP5* fragment of trypsin-treated RRV. Analysis of a Cys-774-->Ser mutant of VP5* showed that, while it was able to form a disulfide bond, a Cys-318-->Ser mutant of VP5* was not. These results indicated that the VP4 component of all rotaviruses, except B223, contains a disulfide bond that links Cys-318 and Cys-380 in the VP5* region of the protein. This bond is located between the trypsin cleavage site and the putative fusion domain of VP4. Because human rotaviruses lack Cys-203 and, hence, unlike many animal rotaviruses cannot possess a disulfide bond in VP8*, it is apparent that VP4 is structurally variable in nature, with human rotaviruses generally containing one disulfide linkage and animal rotaviruses generally containing two such linkages. Considered with the results of anti-VP4 antibody mapping studies, the data suggest that the disulfide bond in VP5* exists within the 2G4 epitope and may be located at the distal end of the VP4 spike on rotavirus particles.

Amino Acid Sequence↗
PubMed1993

CD10/neutral endopeptidase 24.11 regulates fetal lung growth and maturation in utero by potentiating endogenous bombesin-like peptides.

Bombesin-like peptides (BLPs) are mitogens for bronchial epithelial cells and small cell lung carcinomas, and increase fetal lung growth and maturation in utero and in organ cultures. BLPs are hydrolyzed by the enzyme CD10/neutral endopeptidase 24.11 (CD10/NEP) which is expressed in bronchial epithelium and functions to inhibit BLP-mediated growth of small cell lung carcinomas. To determine whether CD10/NEP regulates peptide-mediated lung development, we administered a specific CD10/NEP inhibitor, SCH32615, to fetal mice in utero from gestational days e15-17. Fetal lung tissues were evaluated on e18 for: (a) growth using [3H]thymidine incorporation into nuclear DNA; and (b) maturation using: [3H]-choline incorporation into surfactant phospholipids, electron microscopy for type II pneumocytes, and Northern blot analyses for surfactant apoproteins A, B, and C. Inhibition of CD10/NEP stimulated [3H]thymidine incorporation into DNA (70% above baseline, P < 0.005), [3H]choline incorporation into surfactant phospholipids (38% above baseline, P < 0.005), increased numbers of type II pneumocytes (36% above baseline, P = 0.07), and fivefold higher surfactant protein A transcripts (P < 0.05). CD10/NEP-mediated effects were completely blocked by the specific bombesin receptor antagonist, [D-Phe12, Leu14]bombesin. These observations suggest that CD10/NEP regulates fetal lung growth and maturation mediated by endogenous BLPs.

Animals↗
PubMed1993

A review of the effects of shear and interfacial phenomena on cell viability.

The shear sensitivity of animal and plant cells is a problem often encountered in large-scale cell culture. Such sensitivity varies with different cell lines and the severity of cellular damage may depend on both the magnitude and the duration of the shear stress. In a bioreactor, the shear susceptibility of cells depends on their response to hydrodynamic forces arising from fluid motions of particular scale. Cell damage may be induced by forces in the bulk liquid phase, but fluid motions associated with the gas-liquid interface are especially energetic. The detrimental effects of hydrodynamic forces are abated by the addition of some polymers, such as Pluronic F-68, methylcellulose, or serum; the exact mechanisms of protection are the subject of current research.

Animals↗
PubMed1992

A comparison of cortical strain after cemented and press-fit proximal and distal femoral replacement.

Proximal and distal femoral replacements with intramedullary stems are usually cemented in place but frequently show severe bone remodeling changes in the long term, which can contribute to a loosening process. The remodeling is likely to be associated with stress and strain distribution. This study compared the strain patterns, particularly the maximum principal strains, between cemented and press-fit components in five cadaveric femurs by using a photoelastic coating technique. The specimens were loaded with 2,000 N in the sequences of intact femur and press-fit and cemented stems. On the medial side of the bone for proximal femoral replacements, the strain values for the press-fit stems on the proximal, middle, and distal regions were 73 +/- 11%, 78 +/- 15%, and 80 +/- 15% of normal, respectively, but for the cemented stems they were 53 +/- 15%, 57 +/- 19%, and 60 +/- 20%. The differences were statistically significant (p less than 0.05). On the lateral side, the overall strain values for the press-fit stem were higher than those for the cemented stem, but the differences were not statistically significant (p greater than 0.05). On the medial side of the distal femoral replacement, the strain values for the press-fit stems in the proximal and middle regions were significantly closer to normal than for the cemented stems. Because the press-fit femoral stems (both the proximal and distal replacement) transferred closer to normal strains than the cemented stems, less adverse bone remodeling may be expected, which could be reflected in increased longevity.

Analysis of Variance↗
PubMed1992

Intravascular (catheter) NMR receiver probe: preliminary design analysis and application to canine iliofemoral imaging.

This investigation explores the feasibility of a catheter-based receiver probe for NMR study of arterial walls. Simulations and phantom experiments demonstrate the spatial response of several "inside-out" probe coil designs, including loop, "birdcage," "multipole," "center return," and opposed solenoids. For a target defined by an annulus in a plane perpendicular to B0, the opposed solenoid design provides substantially superior homogeneity to other designs considered. Canine iliofemoral artery images were acquired using a catheter probe in a whole-body, 1.5-T clinical imaging system. In situ (cadaver) images acquired with TE 70, TR 2400, 2-mm slice thickness, and 78 x 78-microns in-plane voxel size in 10-min acquisition times show vessel wall structures identified as intima, internal elastic lamina, media, and adventitia. In vivo images from similar acquistion conditions are much more poorly resolved, presumably due to motion, despite the use of cardiac gating and gradient moment nulling, so the feasibility of obtaining high-resolution in vivo MR images of the arterial wall remains in doubt.

Animals↗
PubMed1992

Signal-to-noise, resolution, and bias function analysis of asymmetric sampling with zero-padded magnitude FT reconstruction.

This report describes NMR image effects due to sampling asymmetry when using zero-padded magnitude FT reconstruction. With this method, the MTF is not flat over the spatial frequency passband, so resolution cannot be accurately described by a single variable such as voxel size. At small to moderate asymmetry, shortened (reduced window duration) asymmetry provides increased S/N and decreased resolution, whereas shifted (constant window duration) asymmetry yields essentially constant S/N with simultaneously increased and decreased resolution. A bias function expression describes image distortion due to sampling in terms separable from the imaged object. The analyses are consistent with previous descriptions of perceived image differences related to data asymmetry.

Algorithms↗
PubMed1992

Mercury distribution in the rat brain after mercury vapor exposure.

Brown Norwegian rats were exposed to mercury vapor at a concentration of approximately 1 mg/m3 for 5 weeks 24 hr/day 7 days a week and 6 hr/day 3 days a week, respectively. The total mercury absorption was calculated to 264 and 35 micrograms per week and 100 g body weight. The mean blood mercury concentration was 0.25 +/- 0.03 and 0.09 +/- 0.01 microgram/g, and the total concentration in the brain was 5.03 +/- 0.73 and 0.71 +/- 0.10 microgram/g tissue, respectively. The mercury distribution in the brains was examined using a method based on chemographic principles. Mercury was found primarily in the neocortex, in the basal nuclei, and in the cerebellar Purkinje cells. This distribution pattern corresponded to the pattern of inorganic mercury described after exposure to methyl mercury. Distribution of mercury after administration of different mercury compounds is discussed.

Administration, Inhalation↗
PubMed1992

CD10/neutral endopeptidase 24.11 in developing human fetal lung. Patterns of expression and modulation of peptide-mediated proliferation.

The cell membrane-associated enzyme CD10/neutral endopeptidase 24.11 (CD10/NEP) functions in multiple organ systems to downregulate responses to peptide hormones. Recently, CD10/NEP was found to hydrolyze bombesin-like peptides (BLP), which are mitogens for normal bronchial epithelial cells and small cell lung carcinomas. Growth of BLP-responsive small cell lung carcinomas was potentiated by CD10/NEP inhibition, implicating CD10/NEP in regulation of BLP-mediated tumor growth. BLP are also likely to participate in normal lung development because high BLP levels are found in fetal lung, and bombesin induces proliferation and maturation of human fetal lung in organ cultures and murine fetal lung in utero. To explore potential roles for CD10/NEP in regulating peptide-mediated human fetal lung development, we have characterized temporal and cellular patterns of CD10/NEP expression and effects of CD10/NEP inhibition in organ cultures. Peak CD10/NEP transcript levels are identified at 11-13 wk gestation by Northern blots and localized to epithelial cells and mesenchyme of developing airways by in situ hybridization. CD10/NEP immunostaining is most intense in undifferentiated airway epithelium. In human fetal lung organ cultures, inhibition of CD10/NEP with either phosphoramidon or SCH32615 increases thymidine incorporation by 166-182% (P < 0.025). The specific BLP receptor antagonist, [Leu13-psi(CH2NH)Leu14]bombesin abolishes these effects on fetal lung growth, suggesting that CD10/NEP modulates BLP-mediated proliferation. CD10/NEP expression in the growing front of airway epithelium and the effects of CD10/NEP inhibitors in lung explants implicate the enzyme in the regulation of peptide-mediated fetal lung growth.

Cell Differentiation↗
PubMed1992

The role of protein synthesis in the decay of phosphoenolpyruvate carboxykinase messenger RNA.

The role of protein synthesis in the control of phosphoenolpyruvate carboxykinase (PEPCK; 4.1.1.32) mRNA turnover was studied in FTO-2B rat hepatoma cells. A previous study demonstrated that incubation of these cells with cAMP prolongs the half-life of the otherwise short-lived PEPCK mRNA. The decay rate of PEPCK mRNA was also slowed in cells incubated with cycloheximide, but not in cells incubated with other translation inhibitors, such as puromycin or pactamycin, even though protein synthesis was inhibited 85-95% by these agents. No correlation was noted between the rate of L-[3H]valine incorporation into cellular proteins and PEPCK mRNA half-life, suggesting that protein synthesis per se is not required for breakdown of the mRNA. Exposure of cells to the translation initiation inhibitor pactamycin together with cycloheximide abolished the "slowing" effect of cycloheximide, and PEPCK mRNA decayed at the same rate as in cells incubated in the presence of pactamycin alone. In contrast, pactamycin did not reverse the effect of cAMP, and the mRNA decayed at the same slow rate in cells incubated in the presence of either (Bu)2cAMP alone or (Bu)2cAMP together with pactamycin. Since pactamycin promotes polysomes dissociation, these results suggest that cAMP enhances the stability of a polysome-free PEPCK mRNA. Furthermore, these results strongly indicate that neither the rapid decay of PEPCK mRNA nor the cAMP-mediated stabilization of the mRNA requires on-going protein synthesis.

Animals↗
PubMed1992

Sleeping through the night: a developmental perspective.

This study examines falling asleep and night waking in human infants during the first 8 months of life. All-night time-lapse video recordings were obtained at 3 weeks and 3 months of age; a Sleep Habits Interview was completed at these ages and repeated at 8 months of age by telephone interview. At the 3-week and 3-month ages, self-report measures of maternal psychologic distress, depression, and self-esteem were also obtained. The data are examined from both cross-sectional (age group) and longitudinal (individual) perspectives. Parent-infant interactions at bedtime and during the middle of the night changed significantly with increasing age. At 3 weeks of age, most infants were put into their cribs for the night already asleep. When they awakened in the middle of the night, they were removed from their cribs. By the time they returned to their cribs, they were again asleep. By 3 months of age, infants who were put into the crib awake at bedtime and allowed to fall asleep on their own were more likely to return to sleep on their own after awakenings later in the night. In contrast, infants who were put into the crib already asleep at the beginning of the night were significantly more likely to be removed from the crib following a subsequent nighttime awakening. Thus, the pattern of bedtime sleep onset was related to sleep onset following an awakening in the middle of the night. This association was present at 8 months as well.(ABSTRACT TRUNCATED AT 250 WORDS)

Child Development↗
PubMed1990

Bombesin increases fetal lung growth and maturation in utero and in organ culture.

Pulmonary neuroendocrine cells (PNECs) in fetuses synthesize gastrin-releasing peptide (GRP, or mammalian bombesin) at high levels, but the role of this hormone in lung development has been obscure. The present study demonstrates that bombesin administered for 2 to 4 d toward the end of gestation in utero led to increased DNA (days 17 and 18) and saturated phosphatidylcholine (SPC) synthesis (day 18) in a dose-dependent fashion in fetal lung. These kinetics coincide with the timing of endogenous GRP gene activation in untreated fetal mouse lung, where GRP mRNA is detectable on day 16 and peaks at day 18. Electron microscopy on in vivo bombesin-treated fetal lung showed an increase in the number of cells containing lamellar bodies on both days 17 and 18, consistent with increased growth and/or maturation of type II cells. In mouse fetal lung organ cultures, the addition of bombesin led to accelerated uptake of [3H]thymidine into DNA, [3H]leucine into protein, and [3H]choline into SPC, indicating that increased growth and maturation may be direct effects. Extending these observations to another species, bombesin was found to induce growth and maturation of human fetal lung in organ culture. A monoclonal antibody to bombesin (2A11) prevented bombesin-induced increases in choline and thymidine incorporation in lung organ cultures and also blocked baseline automaturation of control lung organ cultures in serum-free medium. These data suggest that bombesin, and thus PNECs, play a role in normal lung development.

Animals↗
PubMedDate not supplied

Noise and artifact comparison for Fourier and polynomial phase correction used with Fourier reconstruction of asymmetric data sets.

Two processes of phase correction, Fourier phase mapping and second-order polynomial phase fitting, are compared in combination with four different schemes for Fourier reconstruction of asymmetric data, using one-dimensional simulations and two-dimensional human head data. Polynomial phase correction provides systematically less image noise and is much less affected by localized phase differences caused by object edges and motion.

Artifacts↗
PubMedDate not supplied

Analysis of on- and off-resonance magnetization transfer techniques.

Three methods of performing magnetization transfer (MT) MR imaging are analyzed: (a) off-resonance continuous wave, (b) off-resonance shaped pulses, and (c) on-resonance binomial pulses. With two-pool Bloch-model simulations, signal levels from "MT active" spin systems were calculated, with reference to direct saturation of "MT inactive" systems, allowing calculation of contrast due to MT. Simulations demonstrate several trends with variation of excitation amplitude and offset frequency for the off-resonance methods and with variation of excitation amplitude and pulse shape "order" for binomial pulses. The simulations show that nominally optimized versions of each of these approaches provide essentially equivalent contrast at a given level of applied MT power, contrary to previous claims. Experiments with an MT-inactive phantom, with a whole-body system, show results with off-resonance pulses to be in good agreement with simulations, whereas binomial-pulse experiments show anomalously large direct saturation.

Magnetic Resonance Imaging↗
PubMedDate not supplied

Magnetic resonance imaging of patients wearing a surgical traction halo.

Magnetic resonance images of patients wearing a surgical halo may have unacceptable artifacts if the halo has a conductive loop structure. This study shows that the observed artifacts are predominantly due to eddy currents generated in the halo by switching field gradients, and that these artifacts can be substantially reduced by adjusting the phase encoding direction in MRI sequences so that it is parallel to the axis of the halo.

Artifacts↗