Biomedical subjects

J Hua

Publications and source records attributed to J Hua.

At least 91 records · Page 5Linked to original sources

PubMed1996

Is the coccoid form of Helicobacter pylori viable?

Helicobacter pylori was grown on solid medium for up to 5 weeks. Morphological conversion from spiral to coccoid forms began after 7 days incubation under microaerophilic conditions. Similar to the exponential cultures, the ageing bacteria produced alkaline phosphatase, acid phosphatase, leucine arylamidase and naphthol-AS-beta 1-phosphohydrolase, although there was a reduction in the levels of the latter two enzymes. Unlike the other enzymes, urease was not detected in 5-week-old cultures. By using primers based on urease subunit C and 26 kD protein genes for polymerase chain reaction (PCR) these two important gene fragments remained conserved despite the morphological conversion from spiral to coccoid forms. Furthermore, PCR-based random amplified polymorphic DNA (RAPD) fingerprinting showed similar DNA banding patterns from bacteria of various ages, demonstrating the conservation of the DNA composition despite morphological changes. This study shows that the aging coccoid form of H. pylori, although reportedly non-culturable in vitro, remains genetically unchanged indicating that it is likely to be viable.

Acid Phosphatase↗

PubMed1995

Pathological changes in the Brown Norway rat cerebellum after mercury vapour exposure.

Our previous studies have demonstrated that mercury vapour exposure of Brown Norway rats induced an autoimmune response with development of glomerulonephritis and resulted in mercury deposition in the central nervous system, particularly in the neurons. The aim of this study was to investigate the effect on the central nervous system. A loss of Purkinje cells accompanied by Bergmann glial cell proliferation was found at a brain mercury level of 0.71 micrograms/g and became even more pronounced as the exposure dose increased. At a brain mercury level of 5.0 micrograms/g, a heavy gliosis was present in the brain stem, particularly around the pontine nuclei. In comparison with our previous study, the pathological changes in the brain appeared at the same mercury exposure dose as the glomerulonephritis. However, the location of pathological changes at the mercury level of 0.71 micrograms/g was not completely in accordance with the mercury distribution in the brain, which might be due to the sequence of mercury deposition, its amount or the vulnerability of the various cells classes.

Administration, Inhalation↗

PubMed1995

Ethylene insensitivity conferred by Arabidopsis ERS gene.

ERS (ethylene response sensor), a gene in the Arabidopsis thaliana ethylene hormone-response pathway, was uncovered by cross-hybridization with the Arabidopsis ETR1 gene. The deduced ERS protein has sequence similarity with the amino-terminal domain and putative histidine protein kinase domain of ETR1, but it does not have a receiver domain as found in ETR1. A missense mutation identical to the dominant etr1-4 mutation was introduced into the ERS gene. The altered ERS gene conferred dominant ethylene insensitivity to wild-type Arabidopsis. Double-mutant analysis indicates that ERS acts upstream of the CTR1 protein kinase gene in the ethylene-response pathway.

Amino Acid Sequence↗

PubMed1995

Metabolism and fate of neutral lipids of fetal lung fibroblast origin.

Fetal rat lung fibroblasts characteristically increase their triacylglycerol (TG) stores during development. Both fibroblasts and alveolar type II (TII) cells can synthesize TG de novo, but only fibroblasts can absorb TG from culture medium, and retain the TG in a stable state. When fibroblasts pre-labelled with [3H]triolein are recombined with TII cells in organotypic culture the radiolabel appears in TII cell disaturated phosphatidylcholine (disatPC). When fibroblasts are preloaded with increasing amounts of TG there is a commensurate increase in TII cell disatPC following organotypic culture. Comparison of [3H]triacylglycerol and [14C]glucose incorporation into type II cell phospholipids revealed preferential use of TG for the surface-active phospholipids disatPC (10-fold greater) and phosphatidylglycerol (23-fold greater). These in vitro data suggest that fibroblasts provide lipid substrate for TII cell surfactant phospholipid synthesis.

Animals↗

PubMed1995

Closeness of fit of uncemented stems improves the strain distribution in the femur.

Differences in the cross-sectional shapes of intramedullary stems are expected to affect the strains in the femur, a close proximal fit being proposed as particularly advantageous. We compared uncemented femoral stems of different designs, as well as a cemented stem, using the intact femur as a control. The collarless stems were custom-made, standard asymmetric, symmetric (i.e., neutral), and cemented symmetric. These different types of stem were inserted in sequence into eight cadaver femurs. A photoelastic coating technique was used to give a measure of the shear strains over the bone surface. The mean reductions of maximum shear strains in the upper 30 mm of the proximal-medial regions for the custom, asymmetric, symmetric, and cemented stems were 11, 24, 6, and 75%, respectively. The strains for the symmetric stem were concentrated in the proximal-medial region just below the neck resection line and then decreased rapidly below that level. For the asymmetric stems, the relative strains in the proximal-anterior and proximal-medial regions were dependent on the tightness of fit in the anterior-posterior and medial-lateral directions. The custom stems consistently produced a pattern of strain distribution that, overall, was closer to normal than that of the other stems; extremes at the anterior and medial regions were avoided. The strains down the entire medial side of the cemented stem were significantly less than for the intact femur or for the uncemented stems.

Biomechanical Phenomena↗

PubMed1995

Recognition potential latency and word image degradation.

Ten subjects viewed a stream of meaningless background images in which English words occasionally appeared. A subject indicated detection of a word by lifting the index finger of the preferred hand. He received payment based on his performance. A computer program degraded half of the word images, removing a fraction of the lit pixels and replacing them with others that resembled the background images. The recognition potential (RP) was recorded from occipital electrodes. Degrading the word images increased the peak latency of the RP from 231 to 266 msec. The 35 msec difference was comparable to the 36 msec reaction time (RT) difference. At 395 and 431 msec, RT occurred about 165 msec after the RP peak. The results indicated the RP is closely related to a recognition process. The study did not resolve whether it is concurrent with that process or immediately follows it. The RP's sensitivity to image degradation and its short latency make it unlikely that N2 and P3 are concurrent with the recognition process.

Adult↗

PubMed1995

Progress in identifying clinical relevance of inhibition, stimulation and measurements of poly ADP-ribosylation.

Our laboratory, in collaboration with Oxigene Inc, has been involved in identifying commercially feasible clinical applications of measurement or modulation of ADP-ribosylation as a core technology. For this purpose a pivotal regulatory role for ADP-ribosylation in the repair of DNA lesions leading to cytotoxic as well as mutagenic events has been hypothesized. A new class of DNA repair inhibitors, the N-substituted benzamides, has been identified which can react with radiation to produce reactive intermediates that oxidize thiol amino acids. Their proposed mechanisms of action are two-fold: ie they can interact with radiation: i) to directly enhance DNA damage; and ii) to react with thiols in the zinc finger DNA binding domain of poly ADP-ribosyl transferase to inhibit DNA repair and thereby increase DNA damage. Sensamide, a clinically relevant formulation of metoclopramide which is an N-substituted benzamide, has indicated enhancement of tumor response and survival in patients with inoperable squamous cell carcinoma of the lung when it was administered as a radiosensitizer in a phase I/II trial and compared to historical controls. A mechanism of endogenous regulation of human mononuclear leucocyte ADP-ribosylation has been identified to be HOCl/N-chloramine production via the oxidative burst of phagocytes. HOCl/N-chloramines are potent oxidants of thiol-containing proteins. Quantitative estimation of N-chloramine sensitive plasma thiols has been identified as an effective surrogate measure of leucocyte poly ADPRT.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

Acidic and neutralized metoclopramide formulations sensitize ionizing radiation induced cytotoxicity in a human lung adenocarcinoma xenografted to scid mice.

A neutralized formulation (Neu-Sensamide) of metoclopramide (MCA) has been shown to possess reduced sedative side effects compared with the conventional acidic formulations (Primperan). The acidic formulation of MCA has also been shown to sensitize the effect of ionizing radiation (6-8 Gy) using human squamous cell carcinomas from the head and neck xenografted to nude mice. In the present study, 2 mg MCA/kg body weight 1-3 h before treatment with 1 Gy radiation (single dose) was evaluated in scid mice xenografted with a human lung adenocarcinoma. MCA given alone in acidic or neutralized formulations did not show any effect on tumor growth retardation. However, when combined with radiation, both acidic and neutralized formulations of MCA sensitized the cytotoxic effect of radiation directed against the tumors by increasing tumor doubling time, tumor quadrupling time and specific growth delay, and by decreasing area under growth curve measurements. In addition, there was no statistically significant difference between the two formulations of MCA in the efficacy of sensitizing the cytotoxicity of a single low dose (1 Gy) of radiation.

Adenocarcinoma↗

PubMed1995

Prediction of clinical outcome of THR from migration measurements on standard radiographs. A study of cemented Charnley and Stanmore femoral stems.

We report the theoretical basis of a method to measure axial migration of femoral components of total hip replacements (THR). The use of the top of the greater trochanter and a lateral point on the collar of the stem, allowing for variations of up to 10 degrees rotation of the femur in any direction between successive radiographs, gave a maximum error of 0.37 mm. At a more realistic 5 degrees rotational variation, the error was only 0.13 mm. These data were confirmed in an experimental study using digitisation of points and special software. We also showed that the centre of the femoral head, the stem tip, and the lesser trochanter provided less accurate landmarks. In a second study we digitised a series of radiographs of 51 Charnley and 57 Stanmore THRs; the mean migration rates were found to be identical. We then studied 46 successful stems with a minimum follow-up of eight years and 46 stems which had failed by aseptic loosening at different times. At two years, the successful stems had migrated by a mean of 1.45 +/- 0.68 mm, but the failed cases had a mean migration of 4.32 +/- 2.58 mm (p < 0.0001). Of the successful cases 76% had migrated less than 2 mm, while in the failed group 84% had migrated more than 2 mm. For any particular case migration of more than 2.6 mm at two years had only a 5% chance of continuing success and would therefore merit special follow-up. Only 24% of the eventually successful stems showed migration at the stem-cement interface, but this had happened in every failed stem. We conclude that it would be possible to evaluate a new cemented design of femoral stem over a two-year period by the use of our method and to compare its performance against the reported known standard of the Charnley and Stanmore designs.

Arthritis, Rheumatoid↗

PubMed1995

[Application of laparoscopic technique in splenic surgery].

Five patients of splenic operations had been performed by laparoscopic technique. Patients' ages ranged from 19 to 47, with an average of 31. Four were males and one was female. The procedures included splenectomy 1 patient, deroofing of splenic cyst (1), and hemostasis of splenic injuries (3). The results indicated that all the procedures were successful, and the average length of postoperative stay was 6.5 days. There were no complications. The advantages of laparoscopic splenic surgery include minimal incision, less postoperative pain and rapid recovery. It should be stressed, however, that laparoscopic splenic procedures should be performed cautiously for selected indications only and by surgical endoscopists well trained.

Adult↗

PubMed1994

Parathyroid hormone (PTH) and PTH-related protein stimulate surfactant phospholipid synthesis in rat fetal lung, apparently by a mesenchymal-epithelial mechanism.

We examined the effects of parathyroid hormone (PTH) and PTH-related protein (PTHrP) on rat fetal lung fibroblast and pneumocyte cell signalling. We also studied the effects of PTH and PTHrP on surfactant phospholipid synthesis to determine whether these peptides can modulate pulmonary maturation. Exposure of fibroblasts (gestational days 18-21) to PTH(1-34) or PTHrP(1-34) produced time- and dose-dependent stimulations of cAMP and inositol phosphate accumulation. Maximal stimulation of cAMP accumulation occurred with 1 x 10(-8) M of either peptide. These effects upon cAMP accumulation were competitively inhibited by the PTH antagonist, [Nle8, Nle18, Tyr34]bPTH(3-34)amide. Maximal stimulation of fibroblast inositol phosphates was reached at 1 x 10(-7) M of either peptide. In contrast, PTH and PTHrP at these concentrations produced no changes in cAMP or inositol phosphate metabolism in isolated type II pneumocytes. When pneumocytes were exposed to PTH or PTHrP and pulse-labelled with [methyl-3H]choline chloride, no hormone-stimulated changes in saturated phosphatidylcholine (PC) synthesis were detected. However, PTH and PTHrP stimulated saturated PC synthesis in rat fetal lung explants (gestational day 19-20) by 46% and 106%, respectively. When fibroblasts and pneumocytes were co-cultured, PTH and PTHrP again stimulated saturated PC synthesis by 45% and 73%, respectively. Taken together, these findings suggest that PTH and PTHrP may be endocrine and/or paracrine regulators of fetal lung development.

1,2-Dipalmitoylphosphatidylcholine↗

PubMed1994

Estimation of Bloch model MT spin system parameters from Z-spectral data.

Previous studies have described magnetization transfer (MT) Z-spectra in terms of a two-pool Bloch model, with six spin-system parameters KA, F, T1A, T1B, T2A, and T2B. By simulation, we show that a process including nonlinear constrained optimization can be used to accurately and uniquely estimate spin-system parameters from MT Z-spectra prepared by continuous wave (CW) RF irradiation. Experiments producing Z-spectra by pulsed RF irradiation give substantially different magnetization values, relative to MT acquisitions obtained by CW RF irradiation, at small offset frequencies, with a consequence that only T2B can be uniquely determined. However, several equalities and bounds involving four of the other parameters (KA, F, T1A, and T1B) are derived, which are applicable to pulsed data. These relationships allow calculation of "free pool" magnetization corresponding to complete saturation of the restricted pool, without requiring that this complete saturation be experimentally achieved. MT experimental data from pulsed RF irradiation on boiled egg albumin, obtained using a clinical whole-body MRI system, are analyzed using an optimization algorithm and the derived expressions.

Algorithms↗

PubMed1994

Mercury distribution in cortical areas and fiber systems of the neonatal and maternal adult cerebrum after exposure of pregnant squirrel monkeys to mercury vapor.

Pregnant squirrel monkeys were exposed 5 days/week to mercury vapor at a concentration of 0.5 mg Hg/m3 air for 7 hr/day, or at 1 mg Hg/m3 air for 4 or 7 hr/day. The calculated total mercury absorption ranged between 0.8 and 5.4 mg (range of daily absorption 0.04-0.07 mg). The mercury concentration in the cerebral occipital lobe of the offspring ranged between 0.20 and 0.70 microgram/g tissue, and in the mothers between 0.8 and 2.58 micrograms/g tissue. Mapping of the distribution of mercury in the neocortical layers of the maternal brains revealed that the pyramidal cells contained more visualized mercury than the other neurons. In addition, the mapping disclosed that the deeper the pyramidal cells were situated the more mercury they contained. In the offspring brains, no laminar distribution pattern was found. In the hippocampal formation, the pyramidal cells again contained more mercury than the other neurons. By contrast, the stratum granulosa of the dentate gyrus was always devoid of visualized mercury. The claustrum and the amygdaloid complex always contained mercury. In the fiber systems, the offspring brains contained more mercury than the adult brains. Mercury was found in both glial cells and neurons both in the cortical areas and in the fiber systems.

Air Pollutants↗

PubMed1994

The carboxyl-half of the rotavirus nonstructural protein NS53 (NSP1) is not required for virus replication.

The rotavirus nonstructural protein NS53 (NSVP1), the product of genome segment 5, possesses RNA-binding activity and contains a highly conserved cysteine-rich motif located in the amino-terminal half of the protein. The genome of the bovine rotavirus variant, brvA, lacks a normal segment 5 but includes a novel dsRNA (gene A) of approximately 2600 basepairs (bp) that contains segment 5-specific sequences (F. Hundley, B. Biryahwaho, M. Gow, and U. Desselberger, Virology 143, 88-103, 1985). To gain information about the nature of the rearrangement in gene A and its capacity to encode a protein product, we prepared and sequenced complementary (c)DNA of the gene A RNA. The results showed that gene A is 2693 bp in size and contains a head-to-tail duplication of 1112 bp that originates from the open reading frame (ORF) of gene 5. The duplication begins at nucleotide (nt) 1454, which is 53 nt upstream from the end of the ORF for NS53. Gene A contains a point mutation at nt 808 which results in the presence of a nonsense codon near the middle of the ORF for NS53. Thus the predicted product of gene A is a truncated NS53 of 258 amino acids (aa) (31 kDa), approximately one-half the size of the authentic 491-aa NS53 (58 kDa). Examination of lysates from brvA-infected cells by Western blot assay using an NS53-specific antibody confirmed that the variant encodes only a truncated gene 5 product. Despite the truncation, analysis of the gene A product suggested that it, like full-length NS53, accumulated in association with the cytoskeleton of the infected cell, thus providing evidence that the subcellular localization signal in NS53 resides in the amino terminal half of the protein. Given that brvA is a viable, nondefective mutant, these results demonstrate that the carboxyl-terminal 233 aa of NS53 are not required for rotavirus replication in vitro.

Amino Acid Sequence↗

PubMed1994

Deletion mapping of the rotavirus metalloprotein NS53 (NSP1): the conserved cysteine-rich region is essential for virus-specific RNA binding.

NS53 (NSP1), the gene 5 product of the group A rotaviruses, is a minor nonstructural protein of 486 to 495 amino acids which binds zinc and contains an amino-terminal highly conserved cysteine-rich region that may form one or two zinc fingers. To study the structure-function of the gene 5 product, wild-type and mutant forms of NS53 were produced by using a recombinant baculovirus expression system and a recombinant vaccinia virus/T7 (vTF7-3) expression system. Analysis of the RNA-binding activity of the wild-type NS53 immobilized onto protein A-Sepharose beads with NS53-specific antiserum showed that the protein exhibited specific affinity for all 11 rotavirus mRNAs. The use of short virus-specific RNA probes indicated that NS53 specifically recognizes an element located near the 5' ends of viral mRNAs. Analysis of the RNA-binding activity of deletion mutants of NS53 showed that the RNA-binding domain resides within the first 81 amino acids of the protein and that the highly conserved cysteine-rich region within this region of the protein is essential for the activity. Gel electrophoresis and Western immunoblot analyses of intracellular fractions derived from infected cells revealed that large amounts of NS53 were present in the cytosol and in association with the cytoskeletal matrix. Indirect immunofluorescence analysis of cells programmed to transiently express mutant forms of NS53 using vTF7-3 indicated that the intracellular localization domain resides between amino acids 84 and 176 of NS53. Together, these data show that the RNA-binding domain and the intracellular localization domain lie upstream from the region of NS53 previously determined not to be essential for replication of rotaviruses in cell culture (J. Hua and J. T. Patton, Virology 198:567-576, 1994).

Amino Acid Sequence↗

PubMed1994

Relative motion of hip stems under load. An in vitro study of symmetrical, asymmetrical, and custom asymmetrical designs.

Symmetrical, asymmetrical, and custom asymmetrical stems without collars were manufactured and inserted without cement, in that order, into each of eight femora. Displacement transducers were mounted at different locations to measure relative motion between the stem and the bone. A load was applied to simulate the forces acting on a hip joint in a single-limb stance. A force of 1000 newtons was applied three times in order to seat the prosthesis, and then a cyclical force of 1000 newtons was applied to the head of the prosthesis; readings of relative motion were taken every 500 cycles, until 2500 cycles had been reached. The relative motion diminished as the number of cycles increased, with most of the reduction occurring within the first 500 cycles. The symmetrical stem had the least relative axial motion, with an average of six micrometers, but the other stems also had low values: eleven micrometers for the custom stem, and nineteen micrometers for the asymmetrical stem. In terms of axial rotation, however, the symmetrical stem had several times more motion than did the other two stems. Motion of the stem tip toward or away from the canal in the frontal plane and in the sagittal plane also was higher for the symmetrical stem. Axial migration (permanent sinkage after 2500 cycles) was very similar between all three types of stem. In a torsional loading test, the symmetrical stem showed about ten times more relative rotational motion than did the other two stems. Over-all, the custom stem and the asymmetrical stem had similar results.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1993

Autoimmune glomerulonephritis induced by mercury vapour exposure in the Brown Norway rat.

Subcutaneous injections of mercuric chloride induce an autoimmune glomerulonephritis with both granular and linear IgG deposits along the glomerular capillary wall and proteinuria. This disease is due to a T cell dependent polyclonal B cell activation responsible for production of antibodies against self (glomerular basement membrane, immunoglobulins, DNA, myeloperoxydase) and non self (sheep red blood cells, trinitrophenol (TNP)) components. Increase in serum IgE concentration is the hallmark of this disease. To determine if mercury vapours have pathogenic effects is an important problem of public health. The aim of this study was, first to compare the effects of mercury vapour exposure to those of mercury injections and, second, to compare the effects of high doses to those of low doses of mercury. Two exposure levels were studied corresponding to a mercury absorption of 13.1 mumol/week per kg body wt. and 1.7 mumol/week per kg body wt. during a 5-week period. It will be shown that, whereas the mercury concentration in the kidneys was similar in injected--and vapour exposed--rats, the mercury concentration in blood at the end of the exposure was about twice as high in the injected animals. Blood concentration of mercury was related to dose level but kidney content of mercury was similar in all groups, in spite of a dose difference by a factor of seven between low and high exposure. Mercury vapour and HgCl2 injections both trigger autoimmunity to the same extent and, in both cases the extent of autoimmune manifestations was dose-dependent.

Administration, Inhalation↗

PubMed1993

An RNasin-resistant ribonuclease selective for interleukin 2 mRNA.

Interleukin 2 (IL2) mRNA has a short half-life in the cytoplasm of T lymphocytes, relative to most mRNA. We have discovered a candidate ribonuclease to account for the rapid turnover of IL2 mRNA in the cytosol of the human T lymphocyte cell line Jurkat. In partially purified form, this RNase is about 7 times as active on IL2 as on beta-globin mRNA. Pancreatic RNase, by contrast, does not show a significant preference for IL2 mRNA. Neither 5' capping, nor polyadenylation of the substrate mRNAs affects their degradation by the IL2-selective mRNase, whose activity is optimal in 0.5 mM Mg++ and 100 mM potassium acetate. The mRNase behaves like a protein of molecular weight 60-70,000 on gel chromatography, and is unusual in that it is insensitive to placental RNase inhibitor (RNasin). The mRNase cleaves IL2 mRNA at a small number of sites in the coding region, and IL2 mRNA containing only the coding region and 36 nucleotides of the 3'-noncoding region competes efficiently with full-length IL2 mRNA for the mRNase, whereas beta-globin mRNA does not.

Base Sequence↗