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Biomedical subjects

S Shen

Publications and source records attributed to S Shen.

At least 181 records · Page 10Linked to original sources
PubMed1995

Evidence for linkage of bipolar disorder to chromosome 18 with a parent-of-origin effect.

A susceptibility gene on chromosome 18 and a parent-of-origin effect have been suggested for bipolar affective disorder (BPAD). We have studied 28 nuclear families selected for apparent unilineal transmission of the BPAD phenotype, by using 31 polymorphic markers spanning chromosome 18. Evidence for linkage was tested with affected-sib-pair and LOD score methods under two definitions of the affected phenotype. The affected-sibpair analyses indicated excess allele sharing for markers on 18p within the region reported previously. The greatest sharing was at D18S37: 64% in bipolar and recurrent unipolar (RUP) sib pairs (P = .0006). In addition, excess sharing of the paternally, but not maternally, transmitted alleles was observed at three markers on 18q: at D18S41, 51 bipolar and RUP sib pairs were concordant for paternally transmitted alleles, and 21 pairs were discordant (P = 0004). The evidence for linkage to loci on both 18p and 18q was strongest in the 11 paternal pedigrees, i.e., those in which the father or one of the father's sibs is affected. In these pedigrees, the greatest allele sharing (81%; P = .00002) and the highest LOD score (3.51; phi = 0.0) were observed at D18S41. Our results provide further support for linkage of BPAD to chromosome 18 and the first molecular evidence for a parent-of-origin effect operating in this disorder. The number of loci involved, and their precise location, require further study..

Adolescent↗
PubMed1994

Ectopic expression of Hoxb-8 causes duplication of the ZPA in the forelimb and homeotic transformation of axial structures.

Transgenic embryos were generated carrying a Hoxb-8 transgene under control of the mouse RAR beta 2 promoter, which extends the normal expression domain to more anterior regions of the embryo. These embryos showed mirror-image duplications in the forelimb, analogous to the duplications observed in chick in response to transplantation of a ZPA to the anterior margin of the limb bud. Examination of Sonic hedgehog, Fgf-4, and Hoxd-11 gene expression confirmed that a second ZPA had been generated at the anterior side of the limb bud. Besides other alterations, posterior homeotic transformations of axial structures were observed, involving the first spinal (Froriep's) ganglion and several cervical vertebrae.

Animals↗
PubMed1994

Probing the putative cytochrome P450- and cytochrome c-binding sites on NADPH-cytochrome P450 reductase by anti-peptide antibodies.

Two regions (amino acid residues 110-130 and 204-218) of NADPH-cytochrome P450 reductase (reductase) have been shown to be the putative binding sites for the interaction with cytochrome P450 or cytochrome c. To obtain further insight into the molecular mechanism of protein-protein interaction between these proteins, three anti-peptide antibodies (1A, 2A, and 3A) were generated against the peptides corresponding to these two regions on rat reductase to study the interaction between the reductase and cytochrome P450 or cytochrome c. All three anti-peptide antibodies have high affinity for their peptide antigens on ELISA (titre > 1 x 10(-6) g/L), and they also bind to rat reductase on ELISA under both denatured and native conditions, suggesting that these regions are on the surface of the protein. 1A and 3A also bind to rabbit and human reductase, though 1A binds to human reductase with lower affinity. Antibody 2A does not bind to rabbit or human reductase. Western blot analysis using these anti-peptide antibodies showed similar results. Antibodies 1A and 3A inhibit both cytochrome P4501A1-dependent ethoxycoumarin hydroxylation activity and P4502B1-dependent pentoxyresorufin dealkylation activity, but the inhibition by 1A and 3A was not additive. Antibodies 1A and 3A also have inhibitory effects on the activity of P4501A1-dependent ethoxycoumarin hydroxylation reconstituted with reductase from rabbit and human. However, none of the three anti-peptide antibodies inhibits cytochrome c reduction by rat reductase. These data suggest that reductases from rat, rabbit, and human share similar structure in at least two regions which appear to be on the surface of the protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1994

Comparative toxicity studies of yttrium-90 MX-DTPA and 2-IT-BAD conjugated monoclonal antibody (BrE-3).

BACKGROUND: BrE-3 is monoclonal antibody that has promise for imaging and therapy of human adenocarcinoma. Because of observations in therapeutic trials of yttrium-90 (90Y) escape from radioimmunoconjugates and uptake by the skeleton with resultant bone marrow toxicity, the authors attempted to evaluate the importance of this factor by a comparison of the LD50 in healthy mice treated with 90Y that had been chelated with either of two high affinity chelators, methylbenzyldiethylene-triaminepentaacetic acid (MX-DTPA) or bromoacetamidobenzyl-1,4,7,10-tetraazocyclododecane- N,N',N'',N'''-tetraacetic acid (BAD). METHODS AND RESULTS: Bone marrow hematopoietic toxicity was dose-limiting and the source of death for both chelators. The LD50 for 90Y-BrE-3-MX-DTPA was 220.9 microCi, and that for 90Y-BrE-3-2IT-BAD and was 307.8 microCi. Whole-body autoradiography revealed substantially greater uptake of 90Y in the skeleton when MX-DTPA was used as the chelator. CONCLUSIONS: These observations suggest that 90Y escape to bone is a significant factor in the maximum tolerated dose of radioimmunoconjugate that can be used in therapeutic trials. These results probably underestimate the importance of 90Y escape since 90Y in the skeleton of patients is likely to be more significant than in mice because more of the 90Y energy is absorbed in the marrow of larger species.

Animals↗
PubMed1994

Overview of radiation myelotoxicity secondary to radioimmunotherapy using 131I-Lym-1 as a model.

The radiation dose-limiting toxicity from radioimmunotherapy has been myelotoxicity in the absence of bone marrow reconstitution (transplantation). Myelotoxicity can be assessed directly by biopsy examination of the bone marrow and indirectly by peripheral blood counts. In patients with B-cell malignancies, thrombocytopenia has been the initial and most severe manifestation of 131I-Lym-1 radiation toxicity from treatment. Manifestations of myelotoxicity varied greatly among the patients and from one treatment dose to another in the same patient, suggesting that additional factors were present. There was an increased likelihood of Grade 3-4 hematopoietic toxicity after 131I-Lym-1 treatment if the patient had peripheral blood cell abnormalities before undergoing 131I-Lym-1 treatment. Fractionation of the total 131I-Lym-1 dose was associated with less toxicity. In many patients, myelotoxicity could not be explained by marrow radiation dose (0.36 +/- 0.13 rads per administered mCi) from 131I-Lym-1 in the blood and body alone. Bone marrow examination and 131I-Lym-1 imaging usually provided evidence for additional marrow radiation from 131I-Lym-1-targeting of marrow malignancy and also for residual toxic effects from prior treatment in these patients. Immunohistologic and imaging examination of the bone marrow performed with the intended treatment antibody allowed assessment of extent of marrow malignancy and prediction of degree of myelotoxicity from subsequent treatment. Treatment programs (and protocols) for radioimmunotherapy should incorporate these methods into the decision process. Larger amounts of 131I-Lym-1 can be used in patients selected to have relatively normal peripheral blood cell counts and normocellular bone marrows uninvolved by the malignancy. These observations appear to be relevant to the maximum tolerated dose in radioimmunotherapy for other malignancies as well.

Antibodies, Monoclonal↗
PubMed1994

Ultrasonically assisted electrospray ionization for LC/MS determination of nucleosides from a transfer RNA digest.

The use of an ultrasonic nebulizer to assist electrospray ionization mass spectrometry (ESI-MS) has been described and demonstrated with the analysis of a transfer RNA digest by microcolumn LC. The restricted range of mobile-phase compositions amenable to the electrospray process has traditionally placed a severe limitation on the types of LC applications that can be used with ESI-MS. For this reason, an ultrasonic nebulizer configured for LC has been developed that can generate the fine dispersion of liquid required for ESI-MS from any type of mobile phase. In the case presented here, a transfer RNA was enzymatically digested into its substituent nucleosides, which were then analyzed by microcolumn LC. The required mobile-phase gradient (beginning at 5% methanol) falls outside the solvent range that can be used with conventional electrospray. The ultrasonic nebulizer, however, resolves this problem. The fundamental behavior of the four most common nucleosides (cytidine, adenosine, guanosine, uridine) was studied, and conclusions concerning the effects of solution chemistry were drawn. Specifically, signal from the H+ adducts of these species seems to be strongly dependent on the pKa value. Also, effects from several source operating variables were examined. These included capillary exit voltage, drying and focusing gases, and nebulizer frequency. Performance was found to be consistent over a wide range (0-100% methanol) of mobile-phase compositions. The limit of detection for adenosine injected onto a microcolumn was found to be 100 amol. Finally, nucleosides from as little as 150 fmol of RNA (amount prior to digestion) could be detected.

Chromatography, Liquid↗
PubMed1994

Molecular epidemiology of human rotaviruses in Manipur: genome analysis of rotaviruses of long electropherotype and subgroup I.

In 1987/88 a winter outbreak of infantile gastroenteritis occurred in Manipur, India which was mainly due to rotaviruses of long electropherotype and subgroup (SG) I. The VP7 gene of one of these viruses (M48) has been cloned and sequenced. It was found to be very closely related to the VP7 genes of the G2 serotype human rotaviruses RV-5 and S2. Follow-up epidemiology of this event in Manipur during 1989-1992 yielded mainly rotaviruses of more conventional characteristics (94 isolates of SG II and long electropherotype, and 90 isolates of SG I and short electropherotype), but also 6 isolates of SG I with long electropherotype, indicating that these viruses continue to circulate in the Manipur community. One isolate of short electropherotype was of subgroup II, and one long electropherotype isolate reacted with the group A but not with either the subgroup I or subgroup II monoclonal antibodies.

Amino Acid Sequence↗
PubMed1994

Quantitative bremsstrahlung imaging of yttrium-90 using a Wiener filter.

Yttrium-90 (90Y), a pure beta emitter, is an attractive radionuclide for radioimmunotherapy of cancer. Therapeutic management requires quantitative imaging to measure the pharmacokinetics of the radionuclide in the patient for radiation dosimetric calculations. The bremsstrahlung emissions can be utilized to acquire an image of beta sources using a gamma camera. Quantitation of 90Y by bremsstrahlung imaging is difficult because of poor image quality that results from septal penetration and scatter secondary to the broad bremsstrahlung energies. In this work, quantitative methods for bremsstrahlung imaging of 90Y sources that involved the use of (a) a Wiener filter to deconvolve the septal penetration and scatter while suppressing image noise, and (b) the geometric mean of the conjugate view (GM) and effective point source (EPS) methods to quantify activities were investigated. An abdominal phantom was prepared with 90Y activities in the liver, spleen, tumors, and background volumes that were similar to those observed in patient studies. A twofold improvement in resolution recovery for full width at tenth maximum of the line spread function at 11 cm depth in water was achieved using Wiener restoration. Definition of the organ and tumor edges was greatly enhanced and cross talk between adjacent sources was suppressed after Wiener restoration. These improvements in image quality led to more accurate estimation of organ and tumor activities. Using the optimum attenuation correction method for GM and EPS quantitation of filtered bremsstrahlung images, estimates of individual activities (< or = 17% error) and cumulated activities (< or = 8% error) in all of the sources were accurate except for a tumor of 2 cm diameter. The results of this study provide the basis for a method to quantify beta source distribution and demonstrate the potential use of bremsstrahlung imaging in clinical settings.

Gamma Rays↗
PubMed1994

Rearrangement of the VP6 gene of a group A rotavirus in combination with a point mutation affecting trimer stability.

A group A rotavirus isolated from a lamb with diarrhea in Qinhai province, China, was serially passaged in fetal calf kidney cells. In passage 96, rearrangements of RNA segments 5 and 6 of the viral genome were found. Here we report the nucleotide and predicted amino acid sequences of normal and rearranged RNA 6, coding for the major inner capsid protein VP6. In comparison with the normal gene (N6), the rearranged RNA 6 (R6) contained the normal open reading frame followed by a 473-nucleotide (nt) duplication of the gene beginning 23 nt after the termination codon. The duplicated region starts at nt 768 and runs through to the 3' end of the gene. In accordance with the nucleotide sequence of the rearranged RNA 6, a normal-length VP6 product was found in cells infected with the mutant. However, a single-amino-acid change from proline to glutamine at position 309 slightly affected the electrophoretic mobility of the VP6 monomer of the R6 mutant and reduced the stability of VP6 trimers on gels and at low pH values compared with the normal gene product. The degree of relatedness of VP6 of the Chinese lamb rotavirus Lp14 to those of other group A rotaviruses was determined.

Animals↗
PubMed1994

An optoelectronic registration method as applied to PAF-mediated hydrogen peroxide induced arterial thrombosis.

In the present paper, a standardized method for the induction and registration of platelet thrombi in arterioles (500 microns diameter) of small laboratory animals is described in full detail. Using an optoelectronic analogue computer device, different discriminating parameters characteristic for the thrombotic phenomenon are presented. As the topical application of exogenous PAF-acether induces the generation of endogenous PAF-acether according to previous investigations (Bourgain et al. (1985) Prostaglandins 30, 185) it was deemed interesting to investigate the effect of hydrogen peroxide using the described methodology. It was found that the latter substance not only primes the effect of PAF-acether-induced thrombosis, but also can trigger by itself PAF-acether modulated arterial thrombus formation. Experimental evidence is adduced that these thrombotic phenomena can be most efficiently down regulated by specific PAF-acether antagonists.

Adenosine Diphosphate↗
PubMed1994

Planar gamma camera imaging and quantitation of yttrium-90 bremsstrahlung.

UNLABELLED: Yttrium-90 is a promising radionuclide for radioimmunotherapy of cancer because of its energetic beta emissions. Therapeutic management requires quantitative imaging to assess the pharmacokinetics and radiation dosimetry of the 90Y-labeled antibody. Conventional gamma photon imaging methods cannot be easily applied to imaging of 90Y-bremsstrahlung because of its continuous energy spectrum. METHODS: The sensitivity, resolution and source-to-background signal ratio (S/B) of the detector system for 90Y-bremsstrahlung were investigated for various collimators and energy windows in order to determine optimum conditions for quantitative imaging. After these conditions were determined, the accuracy of quantitation of 90Y activity in an Alderson abdominal phantom was examined. RESULTS: When the energy-window width was increased, the benefit of increased sensitivity outweighed degradation in resolution and S/B ratio until the manufacturer's energy specifications for the collimator were exceeded. Using the same energy window, we improved resolution and S/B for the medium-energy (ME) collimator when compared to the low-energy, all-purpose (LEAP) collimator, and there was little additional improvement using the high-energy (HE) collimator. Camera sensitivity under tissue equivalent conditions was 4.2 times greater for the LEAP and 1.7 times greater for the ME collimators when compared to the HE collimator. Thus, the best, most practical selections were found to be the ME collimator and an energy window of 55-285 keV. When we used these optimal conditions for image acquisition, the estimation of 90Y activity in organs and tumors was within 15% of the true activities. CONCLUSIONS: The results of this study suggest that reasonable accuracy can be achieved in clinical radioimmunotherapy using 90Y-bremsstrahlung quantitation.

Beta Particles↗
PubMed1993

Kinetic study of collagen fixation with polyepoxy fixatives.

A new biomaterial has been developed by fixing native collagen with a polyepoxy compound (PC) fixative. In this study, bovine internal thoracic arteries were fixed with PC under various conditions to help understand the kinetics of the collagen-PC reactions and optimize the fixation process. At predetermined time intervals, small samples were cut from the arteries to determine the quantities of the remaining unreacted amino acids in the collagen. Temperature, concentration, and solution pH were among the key parameters studied. The overall fixation rate was found to be reaction-rate controlled, as the rate of fixation was relatively slow compared with the rate of diffusion of PC. As might be expected, the reaction rate was favored by a higher temperature, concentration, and solution pH. A kinetic model, with a 2.5th reaction order with respect to the reactive functional groups of collagen and a first order with respect to PC, was developed that gave a good fit to the experimental data. Based on this model, the degree of fixation, X, as a function of time, t, is given by (1 - X)-1.5 = 1 + Kt, where K is a constant related to the initial concentrations and the reaction rate constant.

Amino Acids↗
PubMed1993

Role of lysine and arginine residues of cytochrome P450 in the interaction between cytochrome P4502B1 and NADPH-cytochrome P450 reductase.

Chemical modification of cytochrome P450 was used to study the involvement of lysine and arginine residues in the interaction between cytochrome P450 and NADPH-cytochrome P450 reductase. Acetylation of 2.2 and 8.5 mol of lysine/mole of P450 by acetic anhydride led to 38.7 and 95% reductions, respectively, in benzphetamine demethylation activity by NADPH-dependent reconstituted P450/reductase complex, while modification of up to 8.5 mol of lysine/mol of P450 did not inhibit cumene hydroperoxide-supported P450-dependent benzphetamine demethylation. Acetylation of lysine residues by acetic anhydride does not grossly disturb the P450 protein conformation as revealed by absolute, CO-difference and fluorescence spectral studies. Modification of P4502B1 by acetic anhydride did not affect its substrate binding ability either. Lysine residues of P4502B1 putatively involved in the interaction with reductase have been identified by radiolabeling of lysine residues with [14C]acetic anhydride followed by trypsin digestion, HPLC separation, and amino acid microsequencing. Radiolabeled lysines occur at positions 251, 384, 422, 433, and 473. Modification of arginine residues in P4502B1 with phenylglyoxal and 2,3-butanedione seemed to have no significant effect on the benzphetamine demethylation activity of P4502B1 either reconstituted with reductase and NADPH or supported by cumene hydroperoxide. Studies of incorporation of [14C]phenylglyoxal showed no concentration- or time-dependent incorporation of phenylglyoxal into the P4502B1. These results support the hypothesis of a predominant role of lysine residues of P450 in the electrostatic interaction with NADPH-cytochrome P450 reductase.

Acetic Anhydrides↗
PubMed1993

Nucleotide sequences of the VP4 and VP7 genes of a Chinese lamb rotavirus: evidence for a new P type in a G10 type virus.

The VP7 and VP4 genes of a group A rotavirus (Lp14) isolated from a lamb with diarrhea in Qinhai province, China, in 1981 were reverse transcribed, amplified by PCR, cloned, and sequenced. The virus was found to possess a VP7 gene, classifying it as a G10 type, and a VP4 gene which showed only limited homology with any other known VP4 gene, suggesting its classification as a new P type and providing further evidence for the vast diversity of group A rotaviruses.

Amino Acid Sequence↗
PubMed1993

Dominant negative retinoic acid receptor beta.

Induction of the retinoic acid receptor beta 2 (RAR beta 2) gene by retinoic acid (RA) is mediated by a RA response element (RARE), which represents a high affinity binding site for RAR/RXR heterodimers acting at this site as RA-inducible transcription activators. In RA resistant RAC65 cells, RAR beta 2 induction is blocked due to expression of a truncated RAR alpha acting as a dominant negative repressor. Here we show that exogenous expression of RAR but not RXR can restore RA-dependent RAR beta 2 promoter activation in RAC65 cells. Structure-function analysis of hRAR beta 2 mutants in RAC65 cells shows, that the transactivation function required to restore RAR beta 2 promoter activation is dependent on the DNA binding, dimerization and RA-dependent transactivation properties of hRAR beta 2, which are retained in a mutant (beta delta 409) lacking the F domain. By contrast, dominant repression of RA-dependent mRAR beta 2 promoter activation by hRAR beta 2 mutants is independent of the DNA binding or RA-dependent transactivation function but requires a region (residues 204-384) in hRAR beta 2 involved in heterodimerization with RXR. These data extend previous observations on structure-function of RARs and provides tools for studying the role of retinoids and RARs during vertebrate development.

Amino Acid Sequence↗
PubMed1993

[Role of nucleus accumbens in cardiovascular activities and its relationship with opioid peptides in rats].

In this study, electrical stimulation and microinjection in nucleus accumbens in urethane-anesthetized rats were conducted to observe the effect of electrical stimulation of nucleus accumbens on blood pressure and heart rate. The following results were observed: (1) Electrical stimulation of the nucleus accumbens of rat resulted in significant hypotension and bradycardia. (2) Kainic acid microinjected into the nucleus accumbens, could abolish the effects mentioned above. (3) Naloxone administered to the nucleus accumbens could block the cardiovascular inhibitory effect evoked by electrical stimulation of nucleus accumbens. The intra-accumbens injection of mu-receptor agonist, DAGO could also elicit hypotension and bradycardia to an extent comparable to that of the effect due to electrical stimulation, whereas kappa-receptor agonist, U-50 had no such an effect. (4) When the cardiovascular inhibitory effect elicited by electrical stimulation of the nucleus accumbens was observed, the discharge activity in locus ceruleus was also decreased. (5) Bilateral vagotomy could abolish the change in heart rate elicited by the electrical stimulation of nucleus accumbens, but not the hypotension. It is suggested that the mu-opioid receptors of neurons in nucleus accumbens are involved in cardiovascular activity.

Animals↗