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L Zheng

Publications and source records attributed to L Zheng.

At least 253 records · Page 14Linked to original sources
PubMed1996

The interfacial region of dipalmitoylphosphatidylcholine bilayers is perturbed by fusogenic amphipaths.

Several structural methods were used to probe the influence of three fusogenic and four nonfusogenic amphipaths on large, unilamellar dipalmitoylphosphatidylcholine (DPPC) vesicles. For four of these structural measurements there was a correlation observed between the ability of an amphipath to favor poly(ethylene glycol) (PEG)-induced fusion and the structural perturbation reported by each method. First, the fluorescence anisotropy of 1-[4-(trimethylamino)phenyl]-6-phenyhexa-1,3,5-triene (TMA-DPH), which probes the upper region of the bilayer, decreased in the range of PEG concentrations previously found to cause fusion of membranes containing fusogenic amphipaths. For nonfusogenic amphipaths, the anisotropy increased monotonically with PEG concentration. The properties of similar probes that locate in the hydrophobic core of the bilayer showed no correlation with fusogenicity, nor did the properties of probes purported to sense the aqueous surface of the membrane. Second, the frequency of the C=O stretch increased and then decreased dramatically as fusogenic but not nonfusogenic membranes were heated through their phase transition. Third, there was a dramatic increase in the frequency of the C-O-C ester stretch at the membrane order/disorder phase transition for membranes containing fusogenic amphipaths, twice the increase observed for nonfusogenic amphipaths. The spectral characteristics of phosphate, choline, and acyl chain motions showed no such correlation with fusogenicity. Finally, calorimetric measurements showed that low levels of fusogenic amphipaths eliminated the "pretransition" (L beta-->P beta) in DPPC membranes, whereas other amphipaths shifted but did not eliminate this transition. Taken together, these results indicate that fusogenic amphipaths perturb the interface or "backbone" region of the bilayer rather than the hydrophobic core, the headgroup, or the water interface regions of DPPC bilayers.

Calorimetry↗
PubMed1996

The use of 3-methoxymethyl-16 beta, 17 beta-epiestriol-O-cyclic sulfone as the precursor in the synthesis of F-18 16 alpha-fluoroestradiol.

We have prepared 3-methoxymethyl-16 beta, 17 beta-epiestriol-O-cyclic sulfone (1c) and used it as a substrate for the production of F-18 16 alpha-fluoroestradiol, via nucleophilic fluorination with fluoride ion. The compound is straightforward to make from the commercially available epiestriol and is a stable crystalline compound that can be stored for at least a year at room temperature. Reaction with fluorine-18 fluoride provides excellent yields; typically > 90% incorporation of the fluoride is achieved. Partial purification of the labeled product may be accomplished at this stage. Hydrolysis of the methoxymethyl protecting group and ring-opened sulfate occurs rapidly in ethanolic acid solution. In the presence of water the hydrolysis requires more vigorous conditions and additional time but still proceeds to completion. Labeled fluoroestradiol is isolated at the end of a 1-2 h synthesis, depending on the hydrolysis method of 30-45% chemical (decay corrected) yield with respect to fluoride, with a specific activity > 1 Ci per micromole.

Cyclic S-Oxides↗
PubMed1996

Uptake characteristics of loracarbef and cephalexin in the Caco-2 cell culture model: effects of the proton gradient and possible presence of a distinctive second component.

The mechanisms of apical (AP) uptake of cephalexin (CEPH) and loracarbef (LOR) in the absence or presence of an (extemally imposed) proton gradient were determined using well-stirred diffusion chambers that minimize the effects of the unstirred water layer. The results indicated that, compared to AP uptake in the presence of an imposed proton gradient, AP uptake in the absence of an imposed proton gradient had higher K(m) values and lower Jmax values. Furthermore, when inhibition studies were performed in the absence of a proton gradient, only natural peptides were effective, whereas the peptide analogs (e.g., enalapril) were not. In addition to the effects of concentration and competitive inhibitors, the results also indicated that (1) the AP uptake of both drugs was decreased more than 60% by FCCP, regardless of whether the proton gradient was present or absent; (2) effects of protein kinase C promoter were dependent upon the presence of a proton gradient; and (3) AP uptake in the presence of an imposed proton gradient was not affected by feeding restriction, whereas AP uptake in the absence of an imposed proton gradient was. These results showed for the first time that two substrates with similar AP uptake characteristics in the presence of an imposed proton gradient may not share those characteristics in the absence of an imposed proton gradient. Taken together, these results suggest that the AP uptake component that functions in the absence of an imposed proton gradient is distinctly different from the one that functions in the presence of an imposed proton gradient. Data generated from the present study and those in the literature lend support to the hypothesis that this distinctive component represents the second binding site on the AP peptide transporter. However, an alternative hypothesis that there are two AP peptide transporters remains to be disapproved.

Caco-2 Cells↗
PubMed1996

Prevalence of activating ras mutations in morphologically characterized thyroid nodules.

Ras proteins are signal-transducing proteins that share common properties with membrane-anchored G proteins. Mutations at codon 12/13 or codon 61 alter GTP-binding or GTPase activity, respectively. Such activating mutations are present in nearly 30-50% of various malignancies including colon, breast, and lung carcinomas. There are conflicting data regarding the prevalence of ras mutations in the thyroid and their possible pathogenetic role in the different tumor types. To address this question, we examined 45 morphologically characterized thyroid carcinomas, adenomas, and hyperplastic nodules using a highly sensitive single-stranded conformation polymorphism (SSCP) approach combined with DNA-sequencing. DNA from cell lines with known mutations served as controls. A G to A H13 codon substitution replacing an Asp for a Gly residue was detected in 1 papillary carcinoma. Although no H12 or H61 codon substitutions were identified, 2 discrete alterations were identified in codons H17 and 22. No N12/13 codon substitutions were identified. N61 codon substitutions of A to G resulting in a Gly to Arg substitution were detected in 2 papillary carcinomas; the same mutation was also found in one follicular adenoma. Interestingly, K12/13 and K61 ras mutations were not present in any of the tumors examined. These data establish a low prevalence of mutations in all ras gene family members in human thyroid neoplasms. This difference from neoplasms of other organs may explain the relatively indolent biologic behavior of many thyroid tumors and supports an alternate early genetic mutation that is more characteristic of these neoplasms.

Adult↗
PubMed1996

An integrated genetic map of the African human malaria vector mosquito, Anopheles gambiae.

We present a genetic map based on microsatellite polymorphisms for the African human malaria vector, Anopheles gambiae. Polymorphisms in laboratory strains were detected for 89% of the tested microsatellite markers. Genotyping was performed for individual mosquitos from 13 backcross families that included 679 progeny. Three linkage groups were identified, corresponding to the three chromosomes. We added 22 new markers to the existing X chromosome map, for a total of 46 microsatellite markers spanning a distance of 48.9 cM. The second chromosome has 57 and the third 28 microsatellite markers spanning a distance of 72.4 and 93.7 cM, respectively. The overall average distance between markers is 1.6 cM (or 1.1, 1.2, and 3.2 cM for the X, second, and third chromosomes, respectively). In addition to the 131 microsatellite markers, the current map also includes a biochemical selectable markers, Dieldrin resistance (Dl), on the second chromosome and five visible markers, pink-eye (p) and white (w) on the X, collarless (c) and lunate (lu) on the second, and red-eye (r) on the third. The cytogenetic locations on the nurse cell polytene chromosomes have been determined for 47 markers, making this map an integrated tool for cytogenetic, genetic, and molecular analysis.

Animals↗
PubMed1996

Integrated genetic map of Anopheles gambiae: use of RAPD polymorphisms for genetic, cytogenetic and STS landmarks.

Randomly amplified polymorphic DNA (RAPD) markers have been integrated in the genetic and cytogenetic maps of the malaria vector mosquito, Anopheles gambiae. Fifteen of these markers were mapped by recombination, relative to microsatellite markers that had been mapped previously. Thirty-four gel-purified RAPD bands were cloned and sequenced, generating sequence tagged sites (STSs) that can be used as entry points to the A. gambiae genome. Thirty one of these STSs were localized on nurse cell polytene chromosomes through their unique hybridization signal in in situ hybridization experiments. Five STSs map close to the breakpoints of polymorphic inversions, which are notable features of the Anopheles genome. The usefulness and limitations of this integrated mosquito map are discussed.

Animals↗
PubMed1996

Effect of neonatal axoplasmic transport attenuation in the infraorbital nerve on vibrissae-related patterns in the rat's brainstem, thalamus and cortex.

This study evaluated the effects of neonatal attenuation of axoplasmic transport in the infraorbital nerve (ION) on the organization of vibrissae-related patterns in the rat's CNS. Application of colchicine- or vinblastine- impregnated implants to the ION from birth until postnatal day (P)6 to P10 resulted in a 92.4% reduction in the number of trigeminal (V) ganglion cells labelled by application of horseradish peroxidase to the vibrissa pad and a 44.8% decrease in the number of Nissl-stained ganglion cells in the ophthalamic-maxillary portion of the V ganglion. These implants also decreased the number of myelinated fibres in the ION. In normal rats killed on P6-10, there was an average of 10273 +/- 1259 myelinated axons in the nerve. In the animals with colchicine- or vinblastine-treated implants, this value was 3891 +/- 1965. The highest axon count in an experimental animal was 9859. In all animals, axoplasmic transport attenuation resulted in the disappearance of normal vibrissae-related cytochrome oxidase patterns in the brainstem, thalamus and primary somatosensory cortex. Axoplasmic transport attenuation did not result in the disappearance of vibrissae-related ordering of V primary afferent terminal arbors, as demonstrated by anterograde labelling with neurobiotin. These results suggest that some factor conveyed from the periphery of the V ganglion and perhaps on to the brainstem is necessary for the maintenance of vibrissae-related patterns in the thalamus and cortex.

Afferent Pathways↗
PubMed1996

The LIM domain-containing Dbm1 GTPase-activating protein is required for normal cellular morphogenesis in Saccharomyces cerevisiae.

Normal cell growth in the yeast Saccharomyces cerevisiae involves the selection of genetically determined bud sites where most growth is localized. Previous studies have shown that BEM2, which encodes a GTPase-activating protein (GAP) that is specific for the Rho-type GTPase Rho1p in vitro, is required for proper bud site selection and bud emergence. We show here that DBM1, which encodes another putative Rho-type GAP with two tandemly arranged cysteine-rich LIM domains, also is needed for proper bud site selection, as haploid cells lacking Dbm1p bud predominantly in a bipolar, rather than the normal axial, manner. Furthermore, yeast cells lacking both Bem2p and Dbm1p are inviable. The nonaxial budding defect of dbm1 mutants can be rescued partially by overproduction of Bem3p and is exacerbated by its absence. Since Bem3p has previously been shown to function as a GAP for Cdc42p, and also less efficiently for Rho1p, our results suggest that Dbm1p, like Bem2p and Bem3p, may function in vivo as a GAP for Cdc42p and/or Rho1p. Both LIM domains of Dbm1p are essential for its normal function. Point mutations that alter single conserved cysteine residues within either LIM domain result in mutant forms of Dbm1p that can no longer function in bud site selection but instead are capable of rescuing the inviability of bem2 mutants at 35 degrees C.

Amino Acid Sequence↗
PubMed1996

Determination of PO2 and its heterogeneity in single capillaries.

We have applied the phosphorescence lifetime technique (Vanderkooi, J. M., G. Maniara, T. J. Green, and D. F. Wilson. J. Biol. Chem. 262: 5476-5482, 1987) to determine oxygen tension in single capillaries of the hamster retractor muscle. Palladium meso-tetra(4-carboxyphenyl)porphine (10 mg/ml, pH 7.40, bound to bovine serum albumin) was used as the phosphorescent oxygen sensor. Our measurement system consisted of a microscope configured for epi-illumination, a strobe flash lamp, a 430-nm bandpass excitation filter, and a 630-nm cut-on emission filter. A rectangular diaphragm was used to limit the illumination field to 10 microns x 10 microns, and an end-window photomultiplier tube was used to detect the phosphorescence signal, which was then input to an analog-to-digital board in a personal computer. In vitro calibrations were carried out at 37 degrees C on samples flowing through a glass capillary tube (diameter, 300 microns) at four different O2 concentrations (0, 2.5, 5, and 7.5%). In vivo tests were carried out on arterioles, capillaries, and venules of the retractor muscle of anesthetized hamsters. The phosphorescent compound was administered by injection into a jugular vein (20 mg/kg). Phosphorescence decay curves were analyzed by a new model of heterogeneous oxygen distribution in the excitation/emission volume. Mean Po2 values and the local Po2 gradients within the excitation/ emission volume were calculated from phosphorescence life-times obtained from individual decay curves. The time course of Po2 obtained during 0.5-s measurement periods (5 decay curves at 0.1-s intervals) at a given site along a capillary indicated the presence of a gradient in Po2 within the plasma space between and near red blood cells. Similar Po2 gradients were also detected in arterioles and venules. Mean Po2 values for arterioles, capillaries, and venules over the 0.5-s observation period were 27 +/- 5, 14 +/- 2, and 11 +/- 3 (SD) mmHg, respectively. The magnitude of the Po2 gradient in the arterioles, capillaries, and venules was 6 +/- 1, 4 +/- 1, and 2 +/- 1 mmHg/micron, respectively.

Animals↗
PubMed1996

ret/PTC-1, -2, and -3 oncogene rearrangements in human thyroid carcinomas: implications for metastatic potential?

The ret/PTC oncogene is unique to papillary thyroid cancer. Three forms of this oncogene, formed by translocation of three different genes to the tyrosine kinase domain of the ret protooncogene, result in constitutive kinase activation. Correlation with clinical outcome is controversial; ret/PTC-1 has been suggested to predict aggressive behavior. There is no morphological description of ret/PTC-positive tumors. We analyzed 60 thyroid carcinomas for ret/PTC expression to determine correlation with clinical history, disease stage, or tumor morphology. Ribonucleic acid extracted from frozen tissue was reverse transcribed; PCR was performed to amplify ret/PTC-1, 2, and -3. The TPC-1 cell line was the positive control for ret/PTC-1. All tumors were characterized morphologically. Clinical data were collected. The 57 papillary and 3 follicular carcinomas were resected from 44 female patients and 15 males. The average age at diagnosis was 46.2 yr (range. 24-83 yr). Three patients had a history of neck irradiation. At diagnosis, 11 patients had extrathyroidal tumor extension, 20 had lymph node metastases, and 1 had lung metastasis. Thirteen patients had tall cell papillary carcinomas; 3 tumors had focal insular or anaplastic dedifferentiation. The average follow-up was 13.4 months, during which 4 patients had recurrent disease. No deaths occurred. One papillary carcinoma (1.7%) was positive for ret/PTC-1, none was positive for ret/PTC-2, and 2(3.4%) were positive for ret/PTC-3. Although all 3 patients who had tumors containing ret/PTC rearrangements were under the age of 45 yr (range, 26-44 yr) and had small tumors (< 1.2 cm), 2 of these 3 patients presented with lymph node metastases, and the third had lymphatic invasion. ret/PTC oncogene expression did not correlate with radiation history. In summary, ret/PTC oncogene rearrangements were found in 3 of 60 (5%) thyroid carcinomas and were not present in tumors with aggressive morphological features. However, we found ret/PTC rearrangements in young patients (< 45 yr of age) with small thyroid carcinomas showing a predisposition for lymphatic involvement, suggesting a possible role in the development of this subgroup of tumors.

Adult↗
PubMed1996

Identification and characterization of espin, an actin-binding protein localized to the F-actin-rich junctional plaques of Sertoli cell ectoplasmic specializations.

Ectoplasmic specializations are membrane-cytoskeletal assemblages found in Sertoli cells at sites of attachment to elongate spermatids or neighboring Sertoli cells. They are characterized in part by the presence of a unique junctional plaque which contains a narrow layer of parallel actin bundles sandwiched between the Sertoli cell plasma membrane and an affiliated cistern of endoplasmic reticulum. Using a monoclonal antibody, we have identified 'espin,' a novel actin-binding protein localized to ectoplasmic specializations. By immunogold electron microscopy, espin was localized to the parallel actin bundles of ectoplasmic specializations at sites where Sertoli cells contacted the heads of elongate spermatids. The protein was also detected at the sites of ectoplasmic specializations between neighboring Sertoli cells. Espin exhibits an apparent molecular mass of approximately 110 kDa in SDS gels. It is encoded by an approximately 2.9 kb mRNA, which was found to be specific to testis among the 11 rat organs and tissues examined. On the basis of cDNA sequence, espin is predicted to be an 836 amino acid protein which contains 8 ankyrin-like repeats in its N-terminal third, a potential P-loop, two proline-rich peptides and two peptides which contain clusters of multiple glutamates bracketed by arginines, lysines and glutamines in a pattern reminiscent of the repetitive motif found in the protein trichohyalin. The ankyrin-like repeats and a 66 amino acid peptide in the C terminus show significant sequence similarity to proteins encoded by the forked gene of Drosophila. A fusion protein containing the C-terminal 378 amino acids of espin was found to bind with high affinity (Kd = approximately 10 nM) to F-actin in vitro with a stoichiometry of approximately 1 espin per 6 actin monomers. When expressed by transfected NRK fibroblasts, the same C-terminal fragment of espin was observed to decorate actin fibers or cables. On the basis of its structure, localization and properties, we hypothesize that espin is involved in linking actin filaments to each other or to membranes, thereby potentially playing a key role in the organization and function of the ectoplasmic specialization.

Actins↗
PubMed1996

[Intracranial fungal granuloma with CSF space dissemination: a case report].

A 28-year-old male presented with a low grade fever, decreased activity, left hemiparesis and signs of intracranial hypertension. CT showed a moderate hydrocephalus and a large irregular mass in the right temporoparietal region with garland-like enhancement after injection of the contract medium. These findings suggested a malignant brain tumor. MR images demonstrated a mass with low-iso signal intensity on T1 weighted image and low-iso-high mixed intensity on T2, which is like a mosaic pattern. Multiple cerebrospinal fluid space seedings including the wall of the lateral ventricle, the surface of the cerebellum and pons, and the cervical spinal cord were clearly delineated on MR images after Gd-DTPA injection. The large mass was totally removed by craniotomy after ventricle drainage for hydrocephalus. Microscopic examinations showed dense fibrous connective tissue with infiltration of Langhans' giant cells, lymphocytes and fibroblasts around the necrotic centers. These hard components may have been responsible for the low signal intensity on T2-MR images. Many Candida elements were clearly shown with the periodic acid Schiff stain. The diagnosis was that the lesion was an intraparenchymal granuloma due to Candida infection. The patient died on the 8th postoperative day because of brain stem malfunction. Intracranial fungal infection rarely produces a granuloma in the central nervous system. Though it is difficult to diagnose a large irregular mass in the brain, MR images, especially T2 weighted images are useful for the diagnosis of fungal granuloma.

Adult↗
PubMed1996

[Relationship between cellular DNA and expression of epidermal growth factor receptor in pleomorphic adenoma of lacrimal gland].

OBJECTIVE: To obtain the information about whether expression of epidermal growth factor (EGF) receptor is related to the proliferative activity of tumor. METHODS: The expression of EGF receptor and DNA content of pleomorphic adenoma of lacrimal gland in 32 cases were detected by an immunohistochemical avidin biotin peroxidase complex method and determined by an image analysis technique, respectively. The relationship between the expression of EGF receptor and DNA content was analyzed. RESULTS: 10 tumors were stained positively with anti-EGF receptor antibody, the DNA contents of 14 cases were increased, their DNA ploidy distribution pattern showed two or several peaks. Good correlation has been found between the expression of EGF receptor and DNA ploidy distribution pattern, and the DNA distribution pattern of tumor with positive EGF receptor expression showed mainly two or several peaks (P < 0.01). CONCLUSION: The expression of EGF receptor of pleomorphic tumor of lacrimal gland is related to the proliferative activity of tumor cells.

Adenoma, Pleomorphic↗
PubMed1996

[Investigation of infiltrating lymphocytes and immunoglobulin by immunohistochemical method in cheilitis granulomatosa].

Category and distribution of infiltrating lymphocytes and immunoglobulin in 12 cases with cheilitis granulomatosa (CG) and 4 cases with Melkersson-Rosenthal syndrome (MRS) were investigated. The result was that infiltration of lymphocytes in granulomatous type was more severe than that in interstitial inflammtion type. Infiltrators mainly consisted of T helper lymphocytes and immunoglobulin was IgM. It was considered that local immunoreaction was enhanced suggesting that cyto-immunity was a dominant factor with participation of humoral immunity. The result will be helpful to elucidate the pathogeneses and the function of many immuno-factors in the CG. It helps to understand the clinical manifestations and may be a theoretic reference for seeking immuno-modulation for treatment.

Humans↗
PubMed1996

[In vitro culture of neonatal mouse calvaria as a model for study of bone resorption].

We established the in vitro culture system of neonatal mouse calvaria. Bone resorption was determined by the following examinations; (1) the release of calcium from the calvaria into the medium was measured by use of atomic absorption spectrophotometer, and the treated to control ratio was calculated and used as an index of bone resorption; (2) the concentration of osteocalcin in the cultured medium was determined by radioimmunoassay; (3) histological examination. By combined use of the examinations, we were able to objectively determine the bone resorption of cultured calvaria.

Animals↗
PubMed1996

[Effects of interleukin-1 on the differentiation of osteoblasts].

By use of quantitative histochemistry, effects of interleukin 1 (IL1) on the differentiation of osteoblasts derived from neonatal rabbit calvaria were studied. The results showed that IL 1 inhibits the alkaline phosphatase activity in the cultured osteoblasts. Also, both quantitative immunohistochemistry study and radioimmunoassay study showed that IL-1 has the ability to stimulate osteocalcin production and secretion in the cultured osteoblasts. Our results suggest that IL 1 plays a role in the regulation of osteoblastic differentiation, and in the end inhibits the calcification of bone matrix.

Alkaline Phosphatase↗
PubMed1996

[Relationship between expression of P21ras and cellular DNA in pleomorphic adenoma of lacrimal gland].

BACKGROUND: The pleomorphic adenoma is the most frequent tumor of the human lacrimal gland comprising about 50% of the epithelial tumors of this organ. Although being benign, local recurrences can occur when the first removal was incomplete and malignant transformation is also not in frequent. It is well known that many sorts of cellular oncogene products are involved in the initiation, promotion and progression of the human neoplasm. Our purpose was to know whether there is abnormal expression of P21ras in pleomorphic adenoma. METHODS: We have undertaken a study of the expression of P21ras in 5 normal tissues and 32 pleomorphic adenoma of lacrimal gland by immunohistochemical means using the monoclone antibody F-132-62 and the nuclear DNA content in the tumor was assayed by image analysis technique. RESULTS: Normal tissues of lacrimal gland were negative, 12 tumors were stained positively with the antibody. The DNA content of 14 cases of tumor was increased. Their DNA ploidy distribution pattern showed two or several peaks. Good correlation has been found between the expression of P21ras and DNA ploidy distribution pattern, the DNA ploidy distribution pattern of tumor which expressed p21ras showed mainly two or several peaks. P< 0.05. CONCLUSIONS: The result of our studies may suggest that there are increased expression of p21ras in pleomorphic adenoma and the expression of p21ras is related to the promotion and progression of pleomorphic adenoma of lacrimal gland.

Adenoma, Pleomorphic↗