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Biomedical subjects

M G Li

Publications and source records attributed to M G Li.

At least 19 recordsLinked to original sources

Migration pattern of a short femoral neck preserving stem.

UNLABELLED: The CFP stem represents a short collared neck-retaining stem with very proximal metaphyseal anchoring along the calcar combined with up-to-date metallurgy. Despite theoretical advantages, the stability and clinical outcome are unknown. We prospectively measured the migration pattern of this new stem and cup. Twenty-six patients (26 hips) with a mean age of 54 years (range, 40-66 years) underwent THA and were followed for 2 years with radiostereometry, radiographs, and clinical scores. The stem showed some early retroversion (mean, SEM 0.6 degrees, 0.3), but stabilized before 1 year. Subsidence (0.05 mm, 0.06) and varus-valgus tilting (0.03 degrees, 0.01) were low. We observed no bone loss in the calcar region. Factors related to patients, implant design, and implantation did not predict migration patterns. The two-dimensional wear of the ceramic/conventional articulation was 0.09 mm at 2-24 months. The low migration of this short neck preserving stem suggests a favorable long-term outcome but longer followup is needed to substantiate this prediction. This design might become an alternative to standard stems and hip resurfacing. LEVEL OF EVIDENCE: Therapeutic Level IV. See The Guidelines for Authors for a complete description of levels of evidence.

Adult↗

Construction of transgenic Bacillus mucilaginosus strain with improved phytase secretion.

AIMS: To construct a transgenic Bacillus mucilaginosus strain to increase the secretion capability of a wild-type isolate of B. mucilaginosus D4B1 to hydrolyse phytate phosphorus, which can be used as a microbial fertilizer in field application. METHODS AND RESULTS: We constructed a phytase secreting expression vector pSP43 with a mini-Tn5 transposon and a Aspergillus fumigatus phytase expression cassette. The vector pSP43 was successfully transferred into the wild-type B. mucilaginosus using the particle bombardment method, and three transgenic strains with a stable copy of phytase expression cassette integrated into the chromosome of the B. mucilaginosus by Tn5 transposition were selected. The phytase activity of the engineered strains increased 36-46-fold when compared with the wild-type strain of D4B1. CONCLUSIONS: The A. fumigatus phytase gene can be expressed under the direction of p43 promoter in B. mucilaginosus. The expression protein is secreted extracellularly and newly constructed strains showed a high phytase activity. SIGNIFICANCE AND IMPACT OF THE STUDY: A transgenic Bacillus strain by the particle bombardment method was constructed.

6-Phytase↗

The C-terminal module of Chi1 from Aeromonas caviae CB101 has a function in substrate binding and hydrolysis.

The chitinase gene chi1 of Aeromonas caviae CB101 encodes an 865-amino-acid protein (with signal peptide) composed of four domains named from the N-terminal as an all-beta-sheet domain ChiN, a triosephosphate isomerase (TIM) catalytic domain, a function-unknown A region, and a putative chitin-binding domain (ChBD) composed of two repeated sequences. The N-terminal 563-amino-acid segment of Chi1 (Chi1DeltaADeltaChBD) shares 74% identity with ChiA of Serratia marcescens. By the homology modeling method, the three-dimensional (3D) structure of Chi1DeltaADeltaChBD was constructed. It fit the structure of ChiA very well. To understand fully the function of the C-terminal module of Chi1 (from 564 to 865 amino acids), two different C-terminal truncates, Chi1DeltaChBD and Chi1DeltaADeltaChBD, were constructed, based on polymerase chain reaction (PCR). Comparison studies of the substrate binding, hydrolysis capacity, and specificity among Chi1 and its two truncates showed that the C-terminal putative ChBD contributed to the insoluble substrate-protein binding and hydrolysis; the A region did not have any function in the insoluble substrate-protein binding, but it did have a role in the chitin hydrolysis: Deletion of the A region caused the enzyme to lose 30-40% of its activity toward amorphous colloidal chitin and soluble chitin, and around 50% toward p-nitrophenyl (pNP)-chitobiose pNP-chitotriose, and its activity toward low-molecular-weight chitooligomers (GlcNAc)3-6 also dropped, as shown by analysis of its digestion processes. This is the first clear demonstration that a domain or segment without a function in insoluble substrate-chitinase binding has a role in the digestion of a broad range of chitin substrates, including low-molecular-weight chitin oligomers. The reaction mode of Chi1 is also described and discussed.

Aeromonas↗

No relationship between postoperative changes in bone density at the proximal tibia and the migration of the tibial component 2 years after total knee arthroplasty.

The relationship between changes in bone mineral density (BMD) in the proximal tibia and fixation of the tibial component during 2 years postoperatively was investigated in 28 knees. BMD was measured using dual-energy x-ray absorptiometry, and fixation was determined using radiostereometric analysis. BMD decreased at 3 months and returned to baseline level at 24 months, but with large variations on an individual basis. Most of the prosthetic migration occurred within the initial 3 months. The results show that the bone remodeling that occurs during the 2 years after operation has no relation to the migration of the tibial component. The early migration seems to be related more to local activities at the interface rather than to changes in BMD assessed below the interface. The changes in BMD during 2 years reflect the bone remodeling caused by the normalization of alignment after operation and are not related to the implant fixation.

Absorptiometry, Photon↗

Changes in bone mineral density at the proximal tibia after total knee arthroplasty: a 2-year follow-up of 28 knees using dual energy X-ray absorptiometry.

The change in bone mineral density at the proximal tibia during 2 years after total knee arthroplasty was studied in 28 knees (28 patients: 10 men and 18 women; median age: 71 years) with dual energy x-ray absorptiometry. Bone mineral density was measured at the proximal tibia at nine regions of interest below the tibial component within 1 week after the operation (baseline); measurements were repeated at 3, 6, 12, and 24 months. All but one knee was malaligned before the operation, and all but three were corrected to within the normal range of alignment after it. The mean bone mineral density of all nine regions of interest at the proximal tibia temporarily decreased by 13% (p = 0.001) during the initial 3 months, probably due to a general metabolic reaction of the skeleton to the operative trauma combined with the effect of the postoperative immobilization, and then the initial level was regained for as long as 2 years. The overall changes in mean bone mineral density to 2 years were insignificant (p > 0.05); however, a great variation (43.9% decrease to 98.0% increase) was observed on an individual basis. This change over time was significantly associated (R2 = 0.36, p = 0.002) with the level of the baseline bone mineral density, which in turn was partly related (R2 = 0.24, p = 0.009) to the amount of malalignment of the knee before the operation. Knees with high baseline levels (n = 14: 11 with varus and three with valgus alignment) displayed a decrease of 10.0 +/- 14.0% (mean +/- SD, p > 0.05) for as long as 2 years, whereas those with low baseline levels (n = 14: seven with varus and six with valgus alignment and one neutrally aligned) had an increase of 19.1 +/- 38.2% (p = 0.038). In both groups, the mean bone mineral density converged to a level of 0.75-0.95 g/cm2 at 2 years.

Absorptiometry, Photon↗

The effect of the preoperative bone quality on the fixation of the tibial component in total knee arthroplasty.

The relationship between the preoperative level of bone mineral density (BMD) in the proximal tibia and the migration of the tibial component 2 years after total knee arthroplasty was investigated in 28 knees with osteoarthrosis (10 men, 18 women; mean age, 71). Sixteen components were inserted uncemented and 12 were cemented. Mean average BMD measured 10 mm below the joint level was 0.81 g/cm2 (range, 0.15-1.33 g/cm2) and was not influenced by gender, age, weight, or preoperative alignment. Local BMD measured in the medial and lateral condyles was influenced by the preoperative alignment. In knees with uncemented fixation, most of the tibial component migration (ie, subsidence and lift-off) occurred within the first months, and thereafter the implants seemed to stabilize. In the uncemented implants, there was a significant relationship between average BMD and migration (regression analysis with curve-fit estimation). The least migration was seen when average BMD was 0.6 to 1.0 g/cm2. Beyond this range, increased subsidence and lift-off was seen. There was no relationship between BMD and the change in maximum migration between 1 and 2 years postoperatively, however. In knees with cemented fixation, subsidence was initially small but continuously increasing. There were no relationships between BMD and subsidence, lift-off, and maximum migration, indicating that bone-cement can compensate for variations in bone quality, at least in the early period after operation.

Absorptiometry, Photon↗

[Effects of heat and noise environments on lipid peroxidation erythrocyte membrane in pilots].

OBJECTIVE: To investigate the effects of heat and noise environments on lipid peroxidation of erythrocyte membrane in pilots. METHOD: Twenty-four pilots and twenty-one ground personnel (control group) served as the subjects. The pilots performed flying in heat and noise environments. Glutathione peroxidase (GSHpx) and malondialdehyde [correction of malondiadehyde] (MDA) levels in erythrocyte membrane were determined before flying (6:00 a.m.), immediately after flying (12:00 a.m.) and 8 hours after flying (8:00 p.m.) respectively with a spectrophotometer. RESULT: Immediately after flying, GSHpx activity in pilot's erythrocyte membrane decreased significantly as compared with that in control group. Immediately after flying and 8 h after flying, MDA contents in pilots increased significantly as compared with that of control group. CONCLUSION: Heat and noise environments might induce increase of lipid peroxidation reaction and decrease of antioxidant ability.

Adult↗

Filamin is required for ring canal assembly and actin organization during Drosophila oogenesis.

The remodeling of the actin cytoskeleton is essential for cell migration, cell division, and cell morphogenesis. Actin-binding proteins play a pivotal role in reorganizing the actin cytoskeleton in response to signals exchanged between cells. In consequence, actin-binding proteins are increasingly a focus of investigations into effectors of cell signaling and the coordination of cellular behaviors within developmental processes. One of the first actin-binding proteins identified was filamin, or actin-binding protein 280 (ABP280). Filamin is required for cell migration (Cunningham et al. 1992), and mutations in human alpha-filamin (FLN1; Fox et al. 1998) are responsible for impaired migration of cerebral neurons and give rise to periventricular heterotopia, a disorder that leads to epilepsy and vascular disorders, as well as embryonic lethality. We report the identification and characterization of a mutation in Drosophila filamin, the homologue of human alpha-filamin. During oogenesis, filamin is concentrated in the ring canal structures that fortify arrested cleavage furrows and establish cytoplasmic bridges between cells of the germline. The major structural features common to other filamins are conserved in Drosophila filamin. Mutations in Drosophila filamin disrupt actin filament organization and compromise membrane integrity during oocyte development, resulting in female sterility. The genetic and molecular characterization of Drosophila filamin provides the first genetic model system for the analysis of filamin function and regulation during development.

Actins↗

Evidence for cooperative interactions between the two motor domains of cytoplasmic dynein.

Cytoplasmic dynein is a force-transducing ATPase that powers the movement of cellular cargoes along microtubules. Two identical heavy chain polypeptides (> 500 kDa) of the cytoplasmic dynein complex contain motor domains that possess the ATPase and microtubule-binding activities required for force production [1]. It is of great interest to determine whether both heavy chains (DHCs) in the dynein complex are required for progression of the mechanochemical cycle and motility, as observed for other dimeric motors. We have used transgenic constructs to investigate cooperative interactions between the two motor domains of the Drosophila cytoplasmic dynein complex. We show that 138 kDa and 180 kDa amino-terminal fragments of DHC can assemble with full-length DHC to form heterodimeric complexes containing only a single motor domain. The single-headed dynein complexes can bind and hydrolyze ATP, yet do not show the ATP-induced detachment from microtubules that is characteristic of wild-type homodimeric dynein. These results suggest that cooperative interactions between the monomeric units of the dimer are required for efficient ATP-induced detachment of dynein and unidirectional movement along the microtubule.

Adenosine Triphosphate↗

Cerebral cortical blood flow maps are reorganized in MAOB-deficient mice.

Cerebral cortical blood flow (CBF) was measured autoradiographically in conscious mice without the monoamine oxidase B (MAOB) gene (KO, n=11) and the corresponding wild-type animals (WILD, n=11). Subgroups of animals of each genotype received a continuous intravenous infusion over 30 min of phenylethylamine (PEA), an endogenous substrate of MAOB, (8 nmol g-1 min-1 in normal saline at a volume rate of 0.11 microl g-1 min-1) or saline at the same volume rate. Maps of relative CBF distribution showed predominance of midline motor and sensory area CBF in KO mice over WILD mice that received saline. PEA enhanced CBF in lateral frontal and piriform cortex in both KO and WILD mice. These changes may reflect a differential activation due to chronic and acute PEA elevations on motor and olfactory function, as well as on the anxiogenic effects of this amine. In addition to its effects on regional CBF distribution, PEA decreased CBF globally in KO mice (range -31% to -41% decrease from control levels) with a lesser effect in WILD mice. It is concluded that MAOB may normally regulate CBF distribution and its response to blood PEA.

Animals↗

Effects of high temperature and noise on erythrocyte membrane ATPase activity in pilots during flight.

Objective. To determine the effect of heat and noise on erythrocyte membrane ATPase activities in pilots during flying. Method. Twenty-four pilots performing bombing for 3 h (45-53 degrees C, 122-97 dB in the cabin) served as the subjects. 21 ground personnel served as control (27 degrees C in the room). Blood samples were taken from both groups before flying (6:00 a.m.), and immediately (12:00 a.m.) and 8 h (8:00 p.m.) after flying. Na(+)-K+ ATPase, and Ca2(+)-Mg2+ ATPase activities in erythrocyte membrane were determined with colorimetry. Result. The Na(+)-K+ ATPase activity in erythrocyte membrane at 6:00 a.m. in pilots was higher than that in control group at the same time (P<0.01). The Ca2(+)-Mg2+ ATPase activities in erythrocyte membrane at 12:00 a.m. and 8:00 p.m. in pilots were significantly higher, compared with those in control group at the same time (P<0.01). Conclusion. The ATPase values obtained in our study were all within normal range, and the daytime variation of both groups are the same. Exposure of human body to heat and noise for long time may be harmful, the higher ATPase activity is, the more catabolism of ATP will be. ATP exhaustion will lead to Ca2+ overload in erythrocyte thus stiffen the red cell membrane.

Adult↗

Mapping of cerebral metabolic activation in three models of cholinergic convulsions.

Glucose utilization of four cerebral cortex and 35 subcortical regions (CGU) was analyzed in three models of cholinergic seizures induced by the following compounds: 1) soman (pinacolylmethylphosphonofluoridate) an organophosphorus cholinesterase inhibitor, 100 microg/kg SC after pretreatment with pyridostigmine 26 microg/kg IM (n = 6); 2) physostigmine, a carbamate cholinesterase inhibitor, 1.31 mg/kg infused IV over 75 min (n = 6); and 3) pilocarpine, a direct cholinergic agonist, 30 mg/kg SC (n = 6). Physostigmine and pilocarpine were preceded by 3 mmol/kg LiCl IP 20 hrs earlier. Animals injected with saline SC (n = 6) were used as controls. Step-wise discriminant analysis successfully classified 100% of the cases into the four experimental groups with data from only six regions. Pyridostigmine-soman induced the most widespread and greatest increases in CGU. More restricted and lower levels of activation were observed with Li-pilocarpine while Li-physostigmine induced significant increases in CGU only in globus pallidus, entopeduncular nucleus, and substantia nigra. These three regions, which are functionally related, were also activated in the other two models of cholinergic convulsions and may represent the initial step in cholinergic activation of the CNS. Li-pilocarpine failed to activate most of the brainstem and the superior colliculus. All cortical regions were activated by Li-pilocarpine and pyridostigmine-soman, while they were inhibited by Li-physostigmine. This phenomenon may be due in part to the lack of activation with physostigmine of the basal forebrain nuclei (lateral septum, medial septum, vertical and horizontal limbs of the diagonal band, and substantia innominata) resulting in a decreased drive of cortical metabolism.

Animals↗

Cholinesterase inhibition improves blood flow in the ischemic cerebral cortex.

The ability of central cholinesterase inhibition to improve cerebral blood flow in the ischemic brain was tested in Sprague-Dawley rats with tandem occlusion of left middle cerebral and common carotid arteries. Cerebral blood flow was measured with lodo- 14C-antipyrine autoradiography in 170 regions of cerebral cortex. The regional distribution of blood flow was characterized in normal animals by cerebral blood flow maxima in the temporal regions. After 2 h ischemia, minimum cerebral blood flow values were found in the lateral frontal and parietal areas on the left hemisphere, and a new maximum was found in the right hemisphere in an area approximately symmetrical to the ischemic focus. Heptyl-physostigmine (eptastigmine), a carbamate cholinesterase inhibitor with prolonged time of action improved cerebral blood flow in most regions, with the exception of the ischemic core. The drug also enhanced the ischemia-induced rostral shift of cerebral blood flow maxima in the right hemisphere. The effects of eptastigmine were more marked 24 h after ischemia. Discriminant analysis showed that data from only 22 regions was sufficient to achieve 100% accuracy in classifying all cases into the various experimental conditions. The redistribution of cerebral blood flow to the sensorimotor area of the right hemisphere of animals with cerebral ischemia, a phenomenon possibly related to recovery of function, was also enhanced by eptastigmine.

Animals↗

Mapping of cortical metabolic activation in soman-induced convulsions in rats.

The metabolic activation of the cerebral cortex during convulsions induced by the organophosphorus cholinesterase inhibitor soman was studied in detail. Soman was given at a dose equivalent to 0.9 LD50 (100 microgram/kg SC after pretreatment with 26 microgram/kg pyridostigmine, IM, to decrease lethality) to examine separately the metabolic effects of severe acetylcholinesterase inhibition, present always with this dose, and convulsions, present only in some of the animals. Cerebral glucose utilization (CGU) values of cortex divided by CGU of brain stem (nCGU) were calculated for 96 locations in nine coronal slices. Animals injected with pyridostigmine-soman and that developed convulsions (n = 7) showed statistically significant increases of nCGU with regard to animals injected with saline (n = 5) in 33 locations, 27 of which were in a single cluster, with the piriform cortex at its center. Perirhinal cortex, and insular cortex also showed significantly higher nCGU in convulsing rats. Other foci of elevated nCGU were found in frontal and parietal locations. In animals injected with pyridostigmine-soman and that did not develop convulsions (n = 5) in spite of severe cholinesterase inhibition, a single location (piriform cortex) showed significantly higher nCGU than controls. Neuropathology evaluation showed a significant decrease in viable cells only in animals that developed convulsions. This effect correlated with enhanced nCGU. It is concluded that the presence of convulsions, and not exposure to pyridostigmine-soman, determined the pattern of nCGU cortical activation, which correlated closely with the structural changes.

Animals↗

Cholinergic modulation of cerebral cortical blood flow changes induced by trauma.

These experiments tested the role of cholinergic mechanisms in the changes of cerebral cortical blood flow (CBF) induced by brain trauma. CBF was measured with Iodo-14C-antipyrine autoradiography, in 128 cerebral cortex regions of both hemispheres, distributed in eight coronal slices. The effects of a 6.3-mm diameter craniotomy over the left motor-sensory cortex with no weight drop, and of trauma (drop weight of 20 g from 30 cm height on left motor-sensory cortex through a 6.3 mm circular craniotomy) on CBF were studied at 2 and 24 h after the interventions. A group of control animals that received no intervention was also set up. Animals were treated with the cholinesterase inhibitor physostigmine salicylate (3.3 microg/kg/min i.v. infusion started 60 min before CBF measurements), the cholinergic blocker scopolamine hydrobromide (1 mg/kg i.v. pulse, 18 min before CBF measurements), or with the drugs vehicle (saline). A focus of decreased CBF at the site of impact was observed 2 h after trauma, extending caudally as far as the occipital cortex. CBF on the contralateral cerebral cortex was also decreased. Both phenomena reversed partially at 24 h. This spontaneous recovery of CBF was blocked by scopolamine. Physostigmine reversed the decrease in CBF of the traumatized cortex, partially around the contused area and completely in more distant regions. The cerebral cortex contralateral to the trauma showed significantly higher CBF 24 h after trauma when compared to intact controls or craniotomy that peaked at the area symmetrical to the center of trauma. This phenomenon was also enhanced by physostigmine and completely blocked by scopolamine. These results suggest a prominent role of cholinergic mechanisms in the vascular adjustments that accompany cerebral trauma.

Animals↗

The amylose content in rice endosperm is related to the post-transcriptional regulation of the waxy gene.

The waxy (Wx) gene of rice encodes a granule-bound starch synthase (GBSS = waxy protein) required for the synthesis of amylose in endosperm. An analysis of Wx transcripts, Wx protein, and amylose content of 31 rice cultivars revealed that endosperm amylose and Wx protein contents are correlated with the ability of the cultivar to excise intron I from the leader sequence of the Wx transcript. Cultivars with high endosperm amylose content (group I) contain high levels of amylose, Wx protein, and the mature 2.3 kb Wx mRNA. Cultivars with intermediate amylose content (group II) produce substantial amounts of a large 3.3 kb Wx pre-mRNA, with intron I still present, in addition to the mature Wx mRNA, and intermediate levels of Wx protein. Glutinous rice (group III cultivars) contains no amylose, no Wx protein, and no mature Wx mRNA; only the incompletely spliced 3.3 kb Wx pre-mRNA is present in group III cultivars. Based on these results, it is hypothesized that the amylose content of rice endosperm is regulated at the level of Wx transcript processing, and, more specifically, at the stage of intron I excision from the Wx pre-mRNA.

Amino Acid Sequence↗

Root- and shoot-specific responses of individual glutamine synthetase genes of maize to nitrate and ammonium.

The responses of the five cytosolic-type glutamine synthetase (GS1) genes of maize to treatment of hydroponically grown seedlings with 10 mM KNO3 or 10 mM NH4Cl were analyzed. Non-coding 3' gene-specific hybridization probes and radioanalytic imaging were used to quantitate individual gene transcript levels in excised roots and shoots before treatment and at selected times after treatment. Genes GS1-1 and GS1-2 exhibited distinct organ-specific responses to treatment with either nitrogen source. The GS1-1 transcript level increased over three-fold in roots, but changed little if any in shoots. In contrast, the GS1-2 transcript level increased over two-fold in shoots, but decreased in roots after treatment. Increased transcript levels were evident at 4 h after treatment with either nitrogen source, with maximum accumulations present at 8 h after treatment with ammonium and at 10-12 h after treatment with nitrate. The GS1-3 gene transcript level showed little or no change after treatment with either nitrogen source. The GS1-4 gene transcript level remained constant in shoots of treated seedlings, whereas in roots, it exhibited relatively minor, but complex responses to these two nitrogen sources. The GS1-5 gene transcript is present in very small amounts in seedlings, making it difficult to analyze its response to metabolites in young plants. These results provide support for the possibility that different cytosolic GS genes of maize play distinct roles in nitrogen metabolism during plant growth and differentiation.

Blotting, Northern↗

Differential expression of six glutamine synthetase genes in Zea mays.

The maize genome has been shown to contain six glutamine synthetase (GS) genes with at least four different expression patterns. Noncoding 3' gene-specific probes were constructed from all six GS cDNA clones and used to examine transcript levels in selected organs by RNA gel blot hybridization experiments. The transcript of the single putative chloroplastic GS2 gene was found to accumulate primarily in green tissues, whereas the transcripts of the five putative GS1 genes were shown to accumulate preferentially in roots. The specific patterns of transcript accumulation were quite distinct for the five GS1 genes, with the exception of two closely related genes.

Cell Compartmentation↗