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Biomedical subjects

C Chen

Publications and source records attributed to C Chen.

At least 1,225 records · Page 68Linked to original sources
PubMed1986

Pregnancy after human oocyte cryopreservation.

The first successful attempt at deep freezing and thawing of the human oocyte is reported. A twin pregnancy was achieved after insemination and replacement in utero. The procedure involved reduction in size of the oocyte/cumulus-oophorus complex, the addition of the cryoprotectant dimethyl sulphoxide as a one-step procedure, slow cooling between -7 degrees C and -36 degrees C after ice nucleation, and rapid freezing to -196 degrees C before storage in liquid nitrogen. Thawing was achieved rapidly by warming in a 37 degrees C water-bath, followed by dilution of dimethyl sulphoxide as a single step. 80% of forty oocytes showed morphological survival after freezing and thawing. Thirty of these were inseminated; 83% retained their capacity to be fertilised, and 60% proceeded to cleavage division.

Animals↗
PubMed1986

Aflatoxin carryover and clearance from tissues of laying hens.

A feeding trial was conducted to determine the levels of aflatoxins deposited in the tissues of hens fed a diet contaminated with 3310 micrograms AFB1/kg and 1680 micrograms AFB2/kg for 4 wk. At the end of aflatoxin feeding, the livers were pale, enlarged and haemorrhagic and the ovaries were significantly smaller than those from control hens and contained only small ova. Only a small fraction of the aflatoxins consumed was deposited in the tissues, either as the original compounds or as their metabolites, which were widely distributed in all tissues. The highest levels of aflatoxins were detected in the gizzard, kidneys and liver, with average total concentrations of less than 3 micrograms/kg. The lowest residue levels were detected in the breast, blood serum and leg, with breast muscle having a total concentration of less than 0.1 microgram/kg. Two days after removal of the contaminated feed, aflatoxin residues in all tissues had decreased markedly, with no aflatoxins being detected in the heart or spleen. No aflatoxin residues were detected in the breast, leg, gizzard and ovaries of hens killed 8 days after withdrawal, or in the kidneys and blood at 16 days. However, one of seven hens had measurable amounts of AFB2 in the liver 32 days after withdrawal. Although few residues were detected in most tissues after 8 days on the aflatoxin-free diet, variation existed between tissues and between individual hens in the amount of time required to achieve tissue clearance.

Aflatoxins↗
PubMed1986

Fermentative Metabolism of Chlamydomonas reinhardii: III. Photoassimilation of Acetate.

The anaerobic photodissimilation of acetate by Chlamydomonas reinhardii F-60 adapted to a hydrogen metabolism was studied utilizing manometric and isotopic techniques. The rate of photoanaerobic (N(2)) acetate uptake was approximately 20 mumoles per milligram chlorophyll per hour or one-half that of the photoaerobic (air) rate. Under N(2), cells produced 1.7 moles H(2) and 0.8 mole CO(2) per mole of acetate consumed. Gas production and acetate uptake were inhibited by monofluoroacetic acid (MFA), 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU) and by H(2). Acetate uptake was inhibited about 50% by 5% H(2) (95% N(2)). H(2) in the presence of MFA or DCMU stimulated acetate uptake and the result was interpreted to indicate a transition from oxidative to reductive metabolism. Carbon-14 from both [1-(14)C]- and [2-(14)C]acetate was incorporated under N(2) or H(2) into CO(2), lipids, and carbohydrates. The methyl carbon of acetate accumulated principally (75-80%) in the lipid and carbohydrate fractions, whereas the carboxyl carbon contributed isotope primarily to CO(2) (56%) in N(2). The presence of H(2) caused a decrease in carbon lost from the cell as CO(2) and a greater proportion of the acetate was incorporated into lipid. The results support the occurrence of anaerobic and light-dependent citric acid and glyoxylate cycles which affect the conversion of acetate to CO(2) and H(2) prior to its conversion to cellular material.

Journal Article↗
PubMed1986

Early reproductive loss.

Although at least 15% of clinically recognized pregnancies end in spontaneous abortion, a much higher loss varying from 8-78% probably occurs subclinically during very early gestation. This 'early reproductive loss' is largely presumptive and may be caused by factors such as chromosomal abnormalities, infections, endocrine deficiencies, uterine disease, and immunological factors. Little may be offered by way of treatment; but more recently, immunotherapy appears an attractive possibility in selected cases.

Abortion, Spontaneous↗
PubMed1986

Early penetration of human sperm through the vestments of human eggs in vitro.

Penetration of human sperm through the vestments of human oocytes during the first 3 h after insemination was investigated to determine the time taken for sperm capacitation, which precedes the acrosome reaction and fertilization. Twelve oocytes from women who became pregnant by IVF, recovered at laparoscopy after appropriate stimulation, were examined by electron microscopy. Follicular development was controlled by administration of clomiphene citrate, hMG, and hCG. Oocytes were cultured in Whittingham's T6 medium for 7-14 h before insemination and were fixed 1-3 h after insemination with preincubated sperm obtained from fertile men. All oocytes had matured and eight were normally fertilized 2-3 h after insemination. The acrosome reaction had already begun 1 h after insemination when 10-30% of sperm had reacted. Sperm-oocyte membrane fusion occurred 2 h after insemination, and sperm decondensation and pronuclear formation were in progress 3 h after insemination. Variable numbers of sperm (30-60%) had reacted acrosomes after 2-3 h of insemination. Many sperm penetrating the cumulus were intact or had partially reacted acrosomes. Intact, partially and fully reacted sperm were found at the surface of the zona. Intact sperm were bound to the zona by their plasma membranes. Sperm penetrating the zona had reacting or reacted acrosomes exposing their inner acrosome membranes. Those approaching the perivitelline space had a persistent equatorial vestige of the acrosome with intact plasma membrane. The acrosome reaction involved multiple fusions of the sperm plasma and outer acrosome membranes, resulting in vesiculation. This study shows that human sperm could complete capacitation and initiate the acrosome reaction within 1 h of insemination in vitro.

Acrosome↗
PubMed1986

High frequency and nonrandom distribution of alloreactivity in T cell clones selected for recognition of foreign antigen in association with self class II molecules.

Previous studies have demonstrated that a single T cell clone can respond to both a foreign antigen in the context of self major histocompatibility complex (MHC)-encoded molecules (self plus X) and to an allogeneic class I or class II molecule in the absence of antigen (non-self). We have used limiting dilution of T cells obtained from the draining lymph nodes of antigen-primed B10.A mice to establish a large number of T cell clones that recognize either GAT, pigeon cytochrome c, or sheep insulin in association with syngeneic antigen-presenting cells. Sixty-two antigen-specific T cell clones were assayed for their ability to proliferate in response to a panel of nine different allogeneic haplotypes. Of these, 38 (61%) responded to at least one allogeneic haplotype, and 15 of the 38 (39%) responded to more than one allogeneic stimulator. In addition, the patterns of alloreactivity varied with the immunizing antigen. The GAT-specific T cells had at least one responder to every haplotype tested, although H-2u-responsive T cell clones were the most common. In contrast, no pigeon cytochrome c-specific T cells responded to stimulators of the H-2u haplotype, but rather predominantly responded to H-2t4/H-2s and H-2i5/H-2b. Finally, sheep insulin-reactive T cell clones preferentially responded to H-2u stimulators, although stimulation by antigen-presenting cells of the H-2p and H-2q haplotypes was also common. A chi 2 analysis of the data demonstrated that the dependence of the pattern of alloreactivity observed upon the antigen used for immunization was statistically significant (p less than 0.01). The high frequency of alloreactivity found in antigen-specific T cell clones is discussed, as well as the implications that the antigen-dependent skewing of the distribution of alloreactivity have for a one-receptor model vs a two-receptor model of T cell recognition.

Animals↗
PubMed1986

Acute effects of nitrates on left ventricular performance and energetics.

Left ventricular myocardial performance and efficiency were analyzed on the basis of two new energetic parameters in patients with coronary heart disease. The myocardial energy consumed during one cardiac cycle is related on the one hand to the work performed (E1) and on the other to the stress-time integral (E2). E1 was obtained by analysis of the pressure-volume integral divided by the left ventricular muscle mass. E2 was obtained as follows: The stress-time integral was analyzed from pressure-volume data and wall thickness using an ellipsoidal calculation model. In order to transfer the stress-time integral into energy units, the value is multiplied by a constant factor which was obtained experimentally in myothermal studies. In patients with coronary heart disease, a single dose of nitroglycerin (1.6 mg s.l.) reduced the total energy sigma (E1 + E2) from 6.1 mcal/g to 4.7 mcal/g (n = 10; P less than 0.01). The efficiency of myocardial contraction (E1/sigma (E1 + E2] was not changed significantly. These data explain quantitatively the beneficial effect of preload and afterload reduction due to nitroglycerin.

Energy Metabolism↗
PubMed1986

Differences in phosphate metabolite levels in drug-sensitive and -resistant human breast cancer cell lines determined by 31P magnetic resonance spectroscopy.

31P magnetic resonance spectra of perfused human breast cancer cells with the phenotype of pleiotropic drug resistance exhibit striking differences in the levels of phosphate metabolites from the wild-type, drug-sensitive parent cell line. Resistant cells demonstrated elevated levels of phosphocreatine and depressed levels of phosphomonoesters, phosphodiesters, and diphosphodiesters. These differences may reflect significant alterations in the control of bioenergetic metabolism between drug-resistant and -sensitive cells.

Breast Neoplasms↗
PubMed1985

Aversive effects of subcutaneously injected vasopressin in the rat: independence of the ascending dorsal noradrenergic bundle.

The unconditioned stimulus properties of subcutaneously administered arginine vasopressin (AVP) were examined using place and taste conditioning paradigms. Evidence for an aversive effect of the peptide was found, in general agreement with a previous report, although a high dose of AVP (10 micrograms) was required. Also investigated was the possible role of the dorsal noradrenergic bundle (DNAB) in conditioned taste aversion established by AVP. Although the DNAB has been proposed as one of the central pathways through which AVP exerts a facilitatory effect on conditioned behaviour, destruction of this pontine-forebrain projection with 6-hydroxydopamine did not appear to alter the aversiveness of the drug in the present study. These results replicate the finding that AVP can serve as an aversive stimulus but also indicate that the aversive properties of the peptide may prove to be dissociable from its ability to enhance the retention of learned behaviour.

Animals↗
PubMed1985

The circular dichroism properties of phi W-14 DNA containing alpha-putrescinylthymine.

The circular dichroism properties of phi W-14 DNA containing alpha-putrescinylthymine and its acetylated derivative have been examined in a number of aqueous solvents. Native phi W-14 DNA exhibits a B-type CD spectrum whose characteristics do not entirely conform to what would be expected for its GC content (51%). The conformationally sensitive positive band above 260 nm has a rotational strength greater than that normally found in prokaryotic DNAs of comparable GC content, such as Escherichia coli DNA. The rotational strength of this band in the spectrum of the heat-denatured form of phi W-14 DNA, however, is similar to that of heat denatured E. coli DNA. Abolition of the positive charge on the putrescine residues of native phi W-14 DNA by reaction with CH2O or by acetylation reduces the rotational strength to a level appropriate for its GC content. Increases in the electrolyte content of the solvent have the same effect, although the rotational strength of this band in phi W-14 DNA does not become comparable to that of E. coli DNA until 6-7 M LiCl. Titration to pH 10.6 in solvents of modest electrolyte content, however, fails to appreciably affect the CD spectral properties of either native phi W-14 DNA or the derivative in which half of the secondary and all of the primary amino groups have been acetylated. On the basis of these results we have concluded that the enhanced rotational strength of the positive band above 260 nm in the CD spectrum of phi W-14 DNA is due to a conformational difference caused by an ion-pair interaction of the positively charged primary amino groups of putrescine with the phosphate backbone. The CD spectral properties, however, reveal that these differences, averaged over the entire basepair population, appear to be relatively small. The average conformation, at least in dilute aqueous solutions, seems to be an unexceptional B variant with conformational properties which would be more appropriate for a DNA of higher CG content.

Acetylation↗
PubMed1985

Cumene hydroperoxide effected hydroperoxidation by cytochrome P-450.

9-Methylfluorene was found to be oxygenated to 9-hydroperoxy-9-methylfluorene and 9-hydroxy-9-methylfluorene by cytochrome P-450 in the presence of cumene hydroperoxide. Molecular oxygen is required and carbon monoxide is inhibitory. The reaction is inhibited by SKF-525A and metyrapone. Metyrapone and cumene hydroperoxide also retard the conversion of 9-hydroperoxy-9-methylfluorene to 9-hydroxy-9-methylfluorene. The reaction is different from hydroperoxide-supported oxygenation, since the cumene hydroperoxide appears to act as an effector of the enzyme rather than oxygen donor. It is suggested that substrates with stable radicals can be dioxygenated in this manner.

Animals↗
PubMed1985

Hydroperoxidation by cytochrome P-450 oxygenase: metabolism of 9-methylfluorene.

9-Methylfluorene was metabolized by rat liver microsomes to 9-hydroperoxy-9-methylfluorene and 9-hydroxy-9-methylfluorene. The results were confirmed by using a reconstituted cytochrome P-450 oxygenase system purified from phenobarbital-induced rat liver which established its involvement. SKF-525A strongly inhibited the formation of both oxygenation products. Cytochrome P-450 alone brought about the conversion of the hydroperoxide to its alcohol. NADPH augmented the peroxidative reaction, but the presence of NADPH-cytochrome P-450 reductase was without effect. Certain microsomal preparations and reconstituted enzyme yielded little or no detectable amounts of hydroperoxide. This was due to a too rapid conversion of the hydroperoxide to its alcohol. The addition of metyrapone, a compound that inhibited such conversion, resulted in accumulation of 9-hydroperoxy-9-methylfluorene for positive identification. Incubation of 9-methylfluorene with microsomes and NADPH resulted in covalent binding of its metabolite to microsomal proteins. Incubation of 14C-labeled 9-hydroperoxy-9-methylfluorene caused covalent binding of label to proteins, RNA, and DNA.

Animals↗