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Biomedical subjects

L Ma

Publications and source records attributed to L Ma.

At least 181 records · Page 10Linked to original sources
PubMed2000

Inverse treatment planning for Gamma Knife radiosurgery.

An inverse treatment planning system for Gamma Knife radiosurgery has been developed using nonlinear programming techniques. The system optimizes the shot sizes, locations, and weights for Gamma Knife treatments. In the patient's prescription, the user can specify both the maximum number of shots of radiation and a minimum isodose line that must surround the entire treatment volume. After satisfying all of the constraints included in the prescription, the system maximizes the conformity of the dose distribution. This automated approach to treatment planning has been applied retrospectively to a series of patient cases, and each optimized plan has been compared to the corresponding manual plan produced by an experienced user. The results demonstrate that this tool can often improve the tumor dose homogeneity while using fewer shots than were included in the original plan. Therefore, inverse treatment planning should improve both the quality and the efficiency of Gamma Knife treatments.

Algorithms↗
PubMed2000

A dosimetric leaf-setting strategy for shaping radiation fields using a multileaf collimator.

A dosimetric leaf-setting strategy of using multileaf collimators (MLC) for shaping radiation fields has been developed. Existing MLC leaf-setting strategies are all based upon geometric criteria. This new approach, however, matches a prescribed field contour with a MLC using clinically consistent dosimetric criteria. The leaf positions are determined using an iterative optimization algorithm. An empirical dose model was developed to compare the dosimetric-based leaf-setting strategy with the geometric-based leaf-setting strategies. Differences up to half a centimeter in the leaf positions and isodose lines were found between setting the MLC geometrically and setting the MLC dosimetrically. The dosimetric leaf-setting strategy provides the ability to achieve better dose conformation for a clinically desired isodose line. Since the desired isodose line that covers a treatment volume is typically higher than 50% of the maximum dose, the scalloping effects due to the finite leaf width at the leaf edge or 50% isodose lines are much reduced. Another benefit of the dosimetric leaf-setting is that it separates the leaf-setting process from the treatment planning process, and this frees the treatment planning vendors from developing detailed dose models for various existing types and future upgrades of MLC systems.

Algorithms↗
PubMed2000

An investigation of eye lens dose for gamma knife treatments of trigeminal neuralgia.

Stereotactic Gamma Knife radiosurgery has been widely used for treating trigeminal neuralgia (TN). A single large fractional dose of 7000 to 9000 cGy is commonly prescribed as the maximum dose for these treatments. For this reason, if a small percentage of the prescribed dose such as 2-3% scattered to the eye, it could reach or even exceed the tolerance dose of the lens. For several TN cases, we found that the Leksell Gamma Plan system calculates the lens dose about 0.5-2% of the maximum dose independent of the use of eye shielding. These dose values are significantly high and it motivated us to investigate the lens dose for the TN patients treated with stereotactic Gamma Knife radiosurgery. Phantom studies and in vivo dosimetry measurements were carried out for six patients treated at our institution. The average dose to the lens ipsilateral to the treated nerve was measured to be 7.7+/-0.6 cGy. Based on the biological model of Lyman and Emami [Int. J. Radiat. Oncol. Biol. Phys. 21, 109-122 (1991)], the probability of the lens complication (cataract) was determined to be 0.1%. Our findings suggest that few TN patients would develop cataracts after receiving Gamma Knife radiosurgery.

Brain Stem↗
PubMed2000

Identification of G protein-coupled receptor kinase 2 phosphorylation sites responsible for agonist-stimulated delta-opioid receptor phosphorylation.

Agonist-induced receptor phosphorylation is an initial step in opioid receptor desensitization, a molecular mechanism of opioid tolerance and dependence. Our previous research suggested that agonist-induced delta-opioid receptor (DOR) phosphorylation occurs at the receptor carboxyl terminal domain. The current study was carried out to identify the site of DOR phosphorylation during agonist stimulation and the kinases catalyzing this reaction. Truncation (Delta15) or substitutions (T358A, T361A, and S363G single or triple mutants) at the DOR cytoplasmic tail caused 80 to 100% loss of opioid-stimulated receptor phosphorylation, indicating that T358, T361, and S363 all contribute and are cooperatively involved in agonist-stimulated DOR phosphorylation. Coexpression of GRK2 strongly enhanced agonist-stimulated phosphorylation of the wild-type DOR (WT), but Delta15 or mutant DOR (T358A/T361A/S363G) failed to show any detectable phosphorylation under these conditions. These results demonstrate that T358, T361, and S363 are required for agonist-induced and GRK-mediated receptor phosphorylation. Agonist-induced receptor phosphorylation was severely impaired by substitution of either T358 or S363 with aspartic acid residue, but phosphorylation of the T361D mutant was comparable with that of WT. In the presence of exogenously expressed GRK2, phosphorylation levels of T358D and S363D mutants were approximately half of that of WT, whereas significant phosphorylation of the T358/S363 double-point mutant was not detected. These results indicate that both T358 and S363 residues at the DOR carboxyl terminus are capable of serving cooperatively as phosphate acceptor sites of GRK2 in vivo. Taken together, we have demonstrated that agonist-induced opioid receptor phosphorylation occurs exclusively at two phosphate acceptor sites (T358 and S363) of GRK2 at the DOR carboxyl terminus. These results represent the identification of the GRK phosphorylation site on an opioid receptor for the first time and demonstrate that GRK is the prominent kinase responsible for agonist-induced opioid receptor phosphorylation in vivo.

Amino Acid Sequence↗
PubMed2000

Expression and characterization of recombinant human-derived Pneumocystis carinii dihydrofolate reductase.

Dihydrofolate reductase (DHFR) is the target of trimethoprim (TMP), which has been widely used in combination with sulfa drugs for treatment and prophylaxis of Pneumocystis carinii pneumonia. While the rat-derived P. carinii DHFR has been well characterized, kinetic studies of human-derived P. carinii DHFR, which differs from rat-derived P. carinii DHFR by 38% in amino acid sequence, have not been reported to date. Here we report on the expression and kinetic characterization of the recombinant human-derived P. carinii DHFR. The 618-bp coding sequence of the human-derived P. carinii DHFR gene was expressed in Escherichia coli. As determined by sodium dodecyl sulfate-polyacrylamide gel eletrophoresis, the purified enzyme had a molecular mass of 25 kDa, consistent with that predicted from the DNA sequence. Kinetic analysis showed that the K(m) values for dihydrofolate and NADPH were 2.7 +/- 0.3 and 14.0 +/- 4.3 microM, respectively, which are similar to those reported for rat-derived P. carinii DHFR. Inhibition studies revealed that both TMP and pyrimethamine were poor inhibitors of human-derived P. carinii DHFR, with K(i) values of 0.28 +/- 0.08 and 0.065 +/- 0.005 microM, respectively, while trimetrexate and methotrexate were potent inhibitors, with K(i) values of 0.23 +/- 0.03 and 0.016 +/- 0.004 nM, respectively. The availability of purified recombinant enzyme in large quantities should facilitate the identification of antifolate inhibitors with greater potency and higher selectivity for human-derived P. carinii DHFR.

Binding, Competitive↗
PubMed2000

Problems related to determination of MICs of oximino-type expanded-spectrum cephems for Proteus vulgaris.

During in vitro susceptibility testing of clinical isolates of Proteus vulgaris, we noted that the MICs of several expanded-spectrum cephems were much higher in the broth microdilution method than in the agar dilution method (termed the MIC gap phenomenon). Here we investigated the mechanism of the MIC gap phenomenon. Cephems with the MIC gap phenomenon were of the oximino type, such as cefotaxime, cefteram, and cefpodoxime, which serve as good substrates for inducible class A beta-lactamase (CumA) enzymes produced by P. vulgaris; this finding suggests a relationship between the MIC gap phenomenon and CumA. Since peptidoglycan recycling shares a system common to that inducing CumA, we analyzed the mechanism of the MIC gap phenomenon using P. vulgaris B317 and isogenic mutants with mutations in the peptidoglycan recycling and beta-lactamase induction systems. The MIC gap phenomenon was observed in the parent strain B317 but not in B317G (cumG-defective mutant; defective peptidoglycan recycling) and B317R (cumR-defective mutant; defective CumA transcriptional regulator). No beta-lactamase activity was detected in B317G and B317R. beta-Lactamase activity and the MIC gap phenomenon were restored in B317G/pMD301 (strain transcomplemented by a cloned cumG gene) and B317R/pMD501 (strain transcomplemented by a cloned cumR gene). MICs determined by the agar dilution method increased when lower agar concentrations were used. Our results indicated that the mechanism of the MIC gap phenomenon is related to peptidoglycan recycling and CumA induction systems. However, it remains unclear how beta-lactamase induction of P. vulgaris is suppressed on agar plates.

Cephalosporins↗
PubMed2000

The role of polyamines in gastric mucus synthesis inhibited by cigarette smoke or its extract.

BACKGROUND: Cigarette smoking was shown to delay gastric ulcer healing and reduce synthesis of mucus, which is important for gastric ulcer protection and healing. Polyamines are important in these processes. AIMS: To study the effects of cigarette smoking on the synthesis of mucus and to investigate if such an effect is acting by interference with the polyamine pathway. METHODS: Gastric mucosal ornithine decarboxylase activity, mucous secreting layer thickness, and ulcer size were determined after different concentrations of cigarette smoke exposure (0, 2, or 4%) in intact animals and animals with ulcers. Synthesis of mucus and ornithine decarboxylase activity and mRNA expression were also assessed in cigarette smoke extract treated MKN-28 cells. RESULTS: Exposure to cigarette smoke significantly reduced the thickness of the mucous secreting layer and gastric mucosal ornithine decarboxylase activity in animals with or without ulcers. Spermidine not only reversed inhibition of mucus synthesis in both intact and ulcer bearing animals but also reversed the delay in ulcer healing. Cigarette smoke extract significantly reduced mucus synthesis and ornithine decarboxylase activity but not its mRNA expression in MKN-28 cells. The reduction in mucus synthesis was restored by spermidine. CONCLUSIONS: Cigarette smoke and its extract repress mucus synthesis in vivo and in vitro, respectively. Reduction of ornithine decarboxylase activity in gastric mucosa is closely associated with this effect.

Animals↗
PubMed2000

Reduction of EGF is associated with the delay of ulcer healing by cigarette smoking.

Cigarette smoking is associated with peptic ulcer diseases. Smokers have lower levels of salivary epidermal growth factor (EGF) than nonsmokers. We investigated whether reduction of EGF is involved in the delay of gastric ulcer healing by cigarette smoking. Rats with acetic acid-induced ulcers were exposed to cigarette smoke (0, 2, or 4% vol/vol) 1 day after ulcer induction. EGF level was elevated 1 day after ulcer induction in salivary glands and serum, and 4 days after ulcer induction in the gastric mucosa. However, cigarette smoke depressed these beneficial effects and EGF mRNA expression in salivary glands and gastric mucosa. Cigarette smoke delayed gastric ulcer healing and reduced cell proliferation, angiogenesis, and mucus synthesis. Exogenous EGF (10 and 20 microg/kg i.v.) before smoke exposure reversed the adverse effects of cigarette smoke, whereas vascular endothelial growth factor level and nitric oxide synthase activity were unaffected. It is concluded that the detrimental effect of cigarette smoke on ulcer healing is a consequence of reduction of angiogenesis, cell proliferation, and mucus secretion through the depressive action on EGF biosynthesis and its mRNA expression in salivary glands and gastric mucosa.

Animals↗
PubMed2000

Endothelial nitric oxide synthase modulates gastric ulcer healing in rats.

Nitric oxide has been shown to be beneficial for gastric ulcer healing. We determined the relative effects of endothelial and inducible nitric oxide synthases on gastric ulcer healing in rats. Ulcers were induced by serosal application of acetic acid. Ulcer severity, angiogenesis, and nitric oxide synthase expression were assessed 3-10 days later. The effects of inhibitors of nitric oxide synthase were also examined. Inducible nitric oxide synthase mRNA was only detected in ulcerated tissue (maximal at day 3), whereas the endothelial isoform mRNA was detected in normal tissue and increased during ulcer healing. Inducible nitric oxide synthase was expressed in inflammatory cells in the ulcer bed, whereas endothelial nitric oxide synthase was found in the vascular endothelium and in some mucosal cells in both normal and ulcerated tissues. Angiogenesis changed in parallel with endothelial nitric oxide synthase expression. N(6)-(iminoethyl)-L-lysine did not affect angiogenesis or ulcer healing, while N(G)-nitro-L-arginine methyl ester significantly reduced both. In conclusion, endothelial nitric oxide synthase, but not the inducible isoform, plays a significant role in gastric ulcer healing.

Animals↗
PubMed2000

Molecular characterization of the TrkA/NGF receptor minimal enhancer reveals regulation by multiple cis elements to drive embryonic neuron expression.

Neural development relies on stringent regulation of key genes that mediate specialized function. TrkA is primarily expressed in neural crest-derived sensory and sympathetic neurons where it transmits critical survival information. We have identified a 457 base pair sequence upstream of the murine first TrkA coding exon that is conserved in human and in chick, and is sufficient for expression in the correct cells with appropriate timing. Mutation analysis of consensus transcription factor binding domains within the minimal enhancer reveals a complex positive regulation that includes sites required for global expression and sites that are specifically required for DRG, trigeminal or sympathetic expression. These results provide a foundation for identification of the transcriptional machinery that specifies neurotrophin receptor expression.

Animals↗
PubMed2000

Method to determine temperature distribution and intrinsic emissivity of a cavity

A method is presented to determine simultaneously the temperature distribution and the intrinsic emissivities of a cavity surface when radiance distributions along the cavity wall for two wavelengths are given. The intrinsic emissivity and reflection characteristics are assumed not to depend on position on the cavity wall. The intrinsic emissivity and reflection characteristics giving the smallest difference between calculated temperature distributions for the two wavelengths are found. The values found and thus the temperature distributions are verified to be close to the true ones. The method is examined on a cylindrocone by a simulation and applied to radiance temperature distributions measured on a commercially available double cone.

Journal Article↗
PubMed2000

Baculovirus-mediated gene transfer into pancreatic islet cells.

Baculovirus transduction is a gene transfer method that uses a moth cell virus for mammalian cells in culture, which results in a high-level prolonged expression. Here we demonstrate that recombinant baculoviruses can serve as efficient gene transfer vehicles for delivering foreign genes driven by mammalian promoters into human and mouse pancreatic islet cells. Existing methods, such as various transfection and electroporation techniques, either suffer from low efficiency or cause extensive membrane damage. Viral vectors have emerged as an important tool for gene delivery and expression in mammalian cells but suffer from several drawbacks, such as lengthy construction time and expression of viral genes. The baculovirus Autographa californica multiple nuclear polyhedrosis virus is widely used as a vector for expression of foreign genes in insect cells and, more recently, in some mammalian cells. Using several green fluorescent protein- and LacZ-expressing constructs in a cytomegalovirus promoter cassette, we obtained efficient gene expression in primary human and mouse islet cells. There was no impairment of glucose-stimulated intracellular free calcium responses after baculovirus infection. The safety and the relative ease of construction and propagation of the virus makes the baculovirus system a useful tool for facilitating the transfer of foreign genes.

Animals↗
PubMed2000

Ileal bile acid transport regulates bile acid pool, synthesis, and plasma cholesterol levels differently in cholesterol-fed rats and rabbits.

We investigated the effect of ileal bile acid transport on the regulation of classic and alternative bile acid synthesis in cholesterol-fed rats and rabbits. Bile acid pool sizes, fecal bile acid outputs (synthesis rates), and the activities of cholesterol 7alpha-hydroxylase (classic bile acid synthesis) and cholesterol 27-hydroxylase (alternative bile acid synthesis) were related to ileal bile acid transporter expression (ileal apical sodium-dependent bile acid transporter, ASBT). Plasma cholesterol levels rose 2.1-times in rats (98 +/- 19 mg/dl) and 31-times (986 +/- 188 mg/dl) in rabbits. The bile acid pool size remained constant (55 +/- 17 mg vs. 61 +/- 18 mg) in rats but doubled (254 +/- 46 to 533 +/- 53 mg) in rabbits. ASBT protein expression did not change in rats but rose 31% (P < 0.05) in rabbits. Fecal bile acid outputs that reflected bile acid synthesis increased 2- and 2.4-times (P < 0.05) in cholesterol-fed rats and rabbits, respectively. Cholesterol 7alpha-hydroxylase activity rose 33% (24 +/- 2.4 vs. 18 +/- 1.6 pmol/mg/min, P < 0.01) and mRNA levels increased 50% (P < 0.01) in rats but decreased 68% and 79%, respectively, in cholesterol-fed rabbits. Cholesterol 27-hydroxylase activity remained unchanged in rats but rose 62% (P < 0.05) in rabbits. Classic bile acid synthesis (cholesterol 7alpha-hydroxylase) was inhibited in rabbits because an enlarged bile acid pool developed from enhanced ileal bile acid transport. In contrast, in rats, cholesterol 7alpha-hydroxylase was stimulated but the bile acid pool did not enlarge because ASBT did not change. Therefore, although bile acid synthesis was increased via different pathways in rats and rabbits, enhanced ileal bile acid transport was critical for enlarging the bile acid pool size that exerted feedback regulation on cholesterol 7alpha-hydroxylase in rabbits.

Animals↗
PubMed2000

Recurrent lacrimal sac papilloma: case report.

Tumors of the lacrimal sac are rare. Benign papillomas comprise approximately 40% of all neoplasms of the lacrimal drainage system. They often present insidiously with symptoms of dacryostenosis or dacryocystitis. Recurrent bouts of dacryocystitis and nasolacrimal duct obstruction were reported in a 35-year-old man over a period of 13 years. A medial canthal mass was noted in the 6th year after the onset of symptoms. A tumor was discovered incidentally during surgical intervention for presumed dacryostenosis. Surgical removal of the tumor and dacryocystorhinostomy were performed. The histopathologic report turned out to be benign papiloma. Local recurrences occurred during the follow-up period. In addition to surgical excision, we applied cryotherapy and CO2 laser to prevent further recurrence. This case we presented the characteristic recurrence of lacrimal sac papilloma and implied the possibility of tumor occurrence in a patient with recurrent dacryocystitis. We must bear in mind that a recurrent dacryocystitis may be a presentation of a lacrimal sac tumor, because early diagnosis and aggressive treatment can prevent recurrence and result in a cure.

Adult↗
PubMed2000

HER2 promoter controlled specific expression of the reporter gene in ovarian cancer cell line.

OBJECTIVE: Construct a retrovirus vector that expresses the downstream gene specifically in ovarian cancer cells so as to increase the specificity of gene therapy. METHODS: HER2 promoter obtained by PCR was cloned into P(LXSN); mutant GFP cDNA was inserted just downstream of this promoter. Transfect SK-OV3 and MCF-7 cell lines with the new vector, P(LHGSN). RESULTS: Three days after transfection with P(LHGSN), green fluorescence was observed in SK-OV3 cells while not in MCF-7 cells. CONCLUSION: HER2 promoter can control tumor specific expression of the reporter gene in ovarian cancer cell lines. With proper modification, the vector is expected to be useful in tumor specific gene therapy of ovarian cancers.

Female↗
PubMed2000

[Expression and characterization of human vascular endothelial growth factor receptor Flt-1 extracellular domain in Pichia pastoris].

By inserting VEGF receptor Flt-1(1-3 loop) cDNA into Pichia pastoris expression vector pPIC9K containing AOX1 promotor and the sequences of alpha secreting signal peptides, the expression plasmid pPIC9K/Flt-1(1-3) was constructed and transformed into GS115. The multi-copy insert transformants were selected and cultivated in flasks. After 4 days of 1% methanol induction, the expressed Flt-1(1-3) accumulated up to 30% of total proteins in supernatant. The expressed Flt-1(1-3) was further proved with good antigenicity and high specificity by ELISA and Western blot. They can bind to VEGF and inhibit HUVEC proliferation stimulated by VEGF.

Humans↗
PubMed2000

Differential roles of corticotropin-releasing factor receptor subtypes 1 and 2 in opiate withdrawal and in relapse to opiate dependence.

The possible effects on the morphine withdrawal signs of the nonspecific corticotropin-releasing factor (CRF) receptor antagonist alpha-helical CRF, the selective CRF receptor subtype 1 antagonist CP-154,526 and the selective CRF receptor subtype 2 antagonist antisauvagine-30 (AS-30) were investigated in rats. The most withdrawal signs, including jumping, teeth chatter, writhing, shakes, lacrimation, piloerection, irritability and diarrhoea, were attenuated by pretreatment with alpha-helical CRF (10 microg i.c.v.) and CP-154,526 (30 mg/kg i.p.). However, no morphine withdrawal signs except for diarrhea were significantly affected by pretreatment with AS-30 (10 microg, i.c.v.). To investigate the possible role of different CRFR antagonists (alpha-helical CRF, CP-154,526 and AS-30) in relapse to opiate dependence, the 28-day extinction of morphine-conditioned place preference (CPP) was used. The morphine-CPP disappeared following a 28-day extinction and then was reactivated by a single injection of 10 mg/kg morphine. Pretreatment with alpha-helical CRF (10 microg, i.c.v.) and CP-154, 526 (30 mg/kg, i.p.) could significantly block this reactivation of morphine-CPP. In contrast, pretreatment with AS-30 (1 or 10 microg i. c.v.) did not affect this reactivation of morphine-CPP. The present study demonstrated that activation of the CRF receptor is involved in morphine withdrawal signs and relapse to morphine dependence, and that the role of CRF receptor subtypes 1 and 2 in withdrawal and reactivation of morphine dependence is not identical. CRF receptor subtype 1, but not subtype 2, is largely responsible for the action of the CRF system on opiate dependence. These results suggest that the CRF receptor antagonists, particularly the CRF receptor subtype 1 antagonist, might be of some value in the treatment and prevention of drug dependence.

Animals↗