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J Pain

Publications and source records attributed to J Pain.

15 recordsLinked to original sources

Effects of prostaglandin F2alpha treatment on the behavior of pseudopregnant pigs in an extensive environment.

In seminatural environments, prepartum sows leave the herd and construct a maternal nest (a dug out hollow lined with vegetation) prior to the birth of their piglets. The endocrine drives motivating this behavior are not understood, but may involve prostaglandin (PG) F2alpha. This study examined the effect of PGF2alpha treatment on the behavior of pseudopregnant gifts housed in a large enclosure. Pseudopregnancy was induced using 5 mg/ml estradiol valerate/day im from days 11 to 15 of the estrous cycle (first day of estrus = day 0). The gifts' behavior was recorded on a control day, during which no treatment was given, and a test day (= 45.9 +/- 0.42 days of pseudopregnancy) when gilts received either 15 mg PGF2alpha (dinoprost: Lutalyse, Upjohn, Crawley, UK, n = 11) or 0.9% saline (n = 10) im at 11.00 h. PGF2alpha-treated gilts traveled further and were more frequently >10 m from the nearest pig than saline-treated animals. In the hour following injection, PGF2alpha-treated animals also showed increased frequencies of rooting and pawing the ground and stood for longer than saline-treated animals. However, gathering and carrying nest materials were not increased. These results suggest that PGF2alpha, given as a single dose to extensively housed gilts, initiated many, but not all, of the behaviors characteristic of prepartum nest building. The dose and duration of PGF2alpha treatment may have limited the observed behaviors. In addition, environmental feedback is likely to affect the degree to which some nest building behaviors are expressed.

Animals↗

Further characterization of factor VIII-deficient mice created by gene targeting: RNA and protein studies.

Previously we created two strains of factor VIII-deficient mice by insertion of a neo gene into (1) the 3' end of exon 16 and (2) exon 17 of the factor VIII gene. Affected mice of both strains have no plasma factor VIII activity, yet are healthy with no spontaneous bleeding. Factor VIII-deficient females bred with affected males survive pregnancy and delivery. We used reverse transcriptase-polymerase chain reaction of liver RNA to characterize factor VIII mRNA processing. Factor VIII mRNA of the exon 16 knockout strain contains neo sequences plus 17 bp of intron 16 due to use of a cryptic donor site in intron 16. All factor VIII mRNA of the exon 17 knockout strain lacks exon 17 and neo sequences. In skipping exon 17, the intron 16 donor site or a cryptic donor site 46 bp 3' to the intron 16 donor site are used. Thus, factor VIII deficiency in exon 16 knockout mice is due to truncated protein, while in exon 17 knockout mice it is due to either truncated or partially deleted protein. After immunizing exon 16 knockout mice with human recombinant factor VIII, two monoclonal antibodies were obtained that recognize < 100 pg of mouse factor VIII light chain. Assay of cryoprecipitate from the plasma of affected mice failed to show factor VIII light chain.

Animals↗

Ligand-directed immunoaffinity purification and properties of the one-carbon, reduced folate transporter. Interspecies immuno-cross-reactivity and expression of the native transporter in murine and human tumor cells and their transport-altered variants.

Almost complete purification (> 95%) of the 46-kDa murine, one-carbon, reduced folate transporter (RFT) at a recovery of 20% was obtained by ligand-directed immunoaffinity fractionation from transporter overproducing L1210/R83 cells. These cells were labeled with the N-hydroxysuccinimide ester of [3H]aminopterin (AMT), the isolated plasma membrane alkaline washed to remove nonintegral membrane proteins, detergent-solubilized, and RFT-separated on an anti-AMT antibody-protein G-Sepharose column followed by preparative SDS-polyacrylamide gel electrophoresis. Anti-RFT antibody, subsequently derived, differentially blotted (L1210/R83 >> L1210/0) a 46-kDa protein during SDS-polyacrylamide gel electrophoresis of plasma membrane from L1210/R83 and L1210 cells and in L1210/R83 cells after trichloroacetic acid precipitation. In contrast to that reported for human tumor cells, glycosidase treatment of RFT revealed no common N- or O-linked core oligosaccharides associated with this protein. The same 46-kDa protein at different relative levels was revealed in a Western blot of plasma membrane from other murine tumors. Blotting of plasma membrane from methotrexate resistant, transport defective L1210 cell variants exhibited wild-type levels of a less electrophoretically mobile RFT or greater levels of the same 46-kDa RFT which could not be affinity labeled with N-hydroxysuccinimide-[3H]AMT. The same antibody differentially blotted a 83-kDa plasma membrane protein from human HL-60 and CCRF-CEM cells with different levels of reduced folate transport and affinity labeling of RFT, verifying the conserved nature of this protein consistent with earlier functional studies.

Aminopterin↗

Alteration of folate analogue transport inward after induced maturation of HL-60 leukemia cells. Molecular properties of the transporter in an overproducing variant and evidence for down-regulation of its synthesis in maturating cells.

Studies are described examining further the decline in folate analogue influx mediated by the one-carbon reduced-folate transport system in HL-60 cells following induction of maturation by cytodifferentiation agents. To facilitate the investigation of the underlying basis of this phenomenon, we derived a variant (HL-60/LCV) with 4-5-fold elevated influx capacity (Vmax) for folate analogues. A commensurate increase in the putative transporter for this system was documented by affinity labeling of these cells with N-hydroxysuccinimide-[3H]aminopterin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the affinity labeled plasma membrane in HL-60/LCV cells delineated a protein peak at Mr = 75,000-80,000. This was substantially greater than the analogous transporter (Mr = 45,000-47,000) we had delineated (Yang, C.-H., Sirotnak, F.M., and Mines, L.S. (1988) J. Biol. Chem. 263, 9703-9709) with the same methodology in the L1210 cell plasma membrane. In addition, the rate of translocation of the Mr = 75,000-80,000 transporter in HL-60 and HL-60/LCV cells was 2-fold lower than the rate of translocation determined for the Mr = 45,000-47,000 transporter in L1210 cells. During induced maturation of HL-60/LCV cells toward the granulocyte pathway, [3H]methotrexate (MTX) influx capacity and the amount of the affinity labeled transporter decreased rapidly in a parallel fashion. The decrease in [3H]MTX influx and in affinity labeling and in the amount of the Mr = 75,000-80,000 transporter was 5-fold following exposure to 210 mM dimethyl sulfoxide (Me2SO) for 5 days during growth in culture. Moreover, during cycloheximide treatment, the decay in [3H]MTX influx at 37 degrees C and in amount of affinity labeled transporter was the same (t1/2 = 144-155 min) for both control and Me2SO-treated HL-60/LCV cells. These results, which reveal no difference in metabolic turnover for control and Me2SO-treated cells, suggest that the decline in folate analogue influx in HL-60/LCV influx cells is a very early event in the program of differentiation and probably occurs by down-regulation of synthesis of the transporter for the one-carbon reduced-folate transport system.

Affinity Labels↗

Focal nodular hyperplasia of the liver: a link with sickle cell disease?

Focal nodular hyperplasia is a benign liver tumour that is rare in children. We report the second case of a child with sickle cell disease presenting with symptomatic focal nodular hyperplasia. The possible pathogenesis of focal nodular hyperplasia and the association with sickle cell disease are discussed.

Child↗

Surgical options for left-sided large bowel emergencies.

Current choices of operation for left-sided large bowel emergencies have been established by a questionnaire sent to 218 consultant surgeons asking which operation they would perform under varying circumstances for obstructing sigmoid carcinoma and diverticular disease. A 92% response rate was obtained. Hartmann's procedure (with or without a mucus fistula) is the most popular operation for all conditions. Sigmoid colectomy with primary anastomosis is performed by 40% of surgeons for obstructing carcinomas, but less commonly in other situations. On-table lavage is rarely used, and the majority of anastomoses are not protected by a proximal stoma. Subtotal colectomy is very seldom employed, except when caecal perforation results from an obstructing carcinoma. Some surgeons perform a defunctioning colostomy alone even in the presence of a perforation.

Attitude of Health Personnel↗

A comparison of aztreonam/metronidazole and cefotaxime/metronidazole in elective colorectal surgery: antimicrobial prophylaxis must include gram-positive cover.

Aztreonam/metronidazole and cefotaxime/metronidazole were compared in a prospective trial in 154 patients undergoing elective colorectal surgery. Three antibiotic doses were given over 16 hours. A significant excess of wound sepsis in the aztreonam group was seen (23/71 vs 9/70 chi 2 6.60; P less than 0.01). Sepsis was almost invariably due to Gram-positive organisms and, in particular, Staphylococcus aureus.

Aztreonam↗

Altered molecular properties of tubulin in a multidrug-resistant variant of Chinese hamster cells selected for resistance to vinca alkaloids.

The basis for the markedly altered intracellular binding of [3H]vincristine in a multidrug-resistant variant (DC-3F/VCRd-5L) of Chinese hamster lung cells (DC-3F) was investigated. Binding of [3H]vincristine by protein in cytosol derived from each cell type exhibited a differing requirement for GTP in MgCl2 containing buffer of low-ionic strength. Binding of [3H]vincristine occurred to cytosolic protein derived from both variant and parental DC-3F cells, but after removal of GTP, binding only occurred to cytosolic protein from parental cells regardless of the presence of added GTP. Although binding by cytosolic protein from parental DC-3F cells did not require GTP, the addition of 0.1 mM GTP increased by two-fold the rate and extent of binding. When cytosol from variant and parental DC-3F cells was incubated with low concentrations of [3H]vincristine in high-ionic strength buffer and analyzed by molecular-sieve HPLC, most of the protein binding [3H]vincristine in parentally derived cytosol eluted as Mr 110,000-115,000 daltons, corresponding to that for dimeric tubulin. The same binding species was not detected in cytosol derived from variant cells. However, these same fractions derived with both parental and variant cytosols contained tubulin as shown by SDS-PAGE and immunoblotting. A smaller peak of [3H]vincristine binding and an amount of tubulin equal to that found in later fractions were found in the void volume during the same HPLC elution runs with cytosol from both variant and parental DC-3F cells. Evidence was also obtained for differences between parental and variant DC-3F cells in beta-tubulin isoforms following isoelectric focusing and immunoblotting. Parental-cell cytosol contains a single isoform of beta-tubulin. However, in variant cell cytosol the same isoform and, in addition, three more basic isoforms were found. These alterations in [3H]vincristine binding and in isoform compositions of beta-tubulin in variant versus parental DC-3F cells may have importance in regard to vincristine resistance in DC-3F cells.

Animals↗

[In vitro inhibition of tRNA methyltransferases by queen substance, a pheromone of queen honeybees].

The Queen Substance 1, a pheromone of the queen Honeybee Apis mellifica is an in vitro inhibitor of E. coli B tRNA methylations. This activity is not specific of the methylase source, as inhibitions have been observed with preparations from queen honeybee ovaries, Rat liver or a Mouse plasmocytoma 1-adenine methylase. These results, together with preceding ones concerning t, t-farnesyl-acetone 3, are discussed.

Animals↗

[Variation of the 9-keto-2-decenoic acid content as a function of age in virgin queen-bees (Apis mellifica ligustica S.)].

We measured the amount of 9-oxodec-2-enoic acid in virgin Honeybee queens 1 to 50 days old. We chose 4 age ranges: from 1 to 6, from 16 to 20, from 26 to 30, from 40 to 50 days. Queens 16 to 20 days old contain most 9-oxodec-2-enoic acid (average : 1 107 mug). The observed variations in amounts should partly depend on the physiology of attending workers. The authors discuss the mechanisms responsible for the action of 9-oxodec-2-enoic-acid.

Aging↗

[Seasonal variations in the transfer of food labeled with 198Au from the worker to the queen domestic bee].

Food exchanges from a worker giver to a queen receiver were quantified throughout the year. 4 days old workers distribute great amounts of food to queens, about 74% in average, but variations are low. On the other hand workers of unknown age distribute amounts that are less high and varying terms of the season. A parallelism exists between percentage variations of food shared by workers of unknown age and the seasonal cycle of the 9-oxodec-2-enoic acid content of queens.

Age Factors↗

[Radioisotopic demonstration of seasonal variations in food transfer between worker bees].

Food exchanges have been studied during the whole year between workers of unknown age and 4-days old workers. Variations exist between both groups. The mean percentage of shared food is 27% in the group of honeybees of unknown age and 40% in the group of 4-days old workers with respect to the initial intake of the giver. The effect of the season appears among workers of unknown age particularly.

Age Factors↗

Human ecology.

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Air Pollution↗