Determination of embutramide in biological matrices by gas chromatography with nitrogen-phosphorus detection

Embutramide is a general anesthetic having a strong narcotic effect on the central nervous system where it paralyzes the brain center that controls respiration. It is a constituent of T61, a veterinary euthanasia drug. This paper describes a gas chromatographic procedure using nitrogen-phosphorus detection for the determination of embutramide in biological matrices. The drug and the internal standard (ambucetamide) are extracted with dichloromethane under alkaline conditions. The method is linear from 100 to 3000 ng/ml. The within-day and day-to-day coefficients of variation range from 5.1 to 5.7% and from 9.1 to 10.0%, respectively. The recovery is above 80% while the minimum detectable level under the conditions described is 40 ng/ml analyzing a 1-ml or a 1-g aliquot of a sample (blood or tissue). The method is also applied to different samples from dogs euthanized with T61.     

Molecular genetic analysis of the pullulanase B gene of Bacillus acidopullulyticus

In this study we describe immunotoxins prepared with different CD2 monoclonal antibodies (mAbs) and a ribosome-inactivating protein, saporin. The CD2 immunotoxins were tested on different models. Anti-CD2-saporin conjugates inhibited protein synthesis by a neoplastic CD2+ cell line (SKW-3) and by an interleukin 2 dependent polyclonal CD2+ lymphoid cell culture (T lymphoblasts), with IC50s ranging from 10(-13) M to 10(-11) M (as saporin). Similar results were obtained with proliferation inhibition tests (3H-thymidine incorporation) on phytohaemagglutinin (PHA) driven lymphoid cultures and on mixed lymphocyte culture activated lymphocytes. Moreover a CD2-ricin A chain conjugate was less effective than an analogous immunotoxin containing the same CD2 mAb and saporin in inhibiting lymphocyte proliferation induced by PHA (IC50 approximately 10(-9) M as ricin A chain versus 10(-12) M as saporin). The conjugates were not toxic on bone marrow stem cells. These results suggest that CD2-saporin immunotoxins could represent an effective tool for CD2+ lymphomas or leukaemias, and for T-dependent immune disorders, such as transplanted organ rejection and graft-versus-host disease.     

Gastrointestinal complications after aortocoronary bypass

Under analysis are gastrointestinal complications (hemorrhage, perforation of the duodenum, acute pancreatitis) which were observed in 10 of 177 patients after aortocoronary bypass. Risk factors of these complications, their specific diagnosis, treatment and prophylactics are discussed.     

The effect of 5-bromo-2'-deoxyuridine on the lifespan and behavior of Drosophila melanogaster

In this study, we have examined the effect of 5-bromo-2'-deoxyuridine incorporated into DNA of Drosophila nerve cells after incubation of larvae in the analog-containing medium on the duration of lifespan and behavior (photoactivity) of adult flies. 5-Bromo-2'-deoxyuridine incorporated into DNA decreases the lifespan of adult animals. In contrast to the control, the curves describing the probability of death become bimodal at higher doses of the analog (above 35 (g/ml). As the dose of 5-bromo-2'-deoxyuridine decreases, photoactivity of the flies diminishes, and the distribution into fractions with different activity becomes broader. The data obtained provide evidence that the modification of DNA with 5-bromo-2'-deoxyuridine drastically changes the expression of tissue-specific genes in nerve ganglia of Drosophila and at the same time diminishes the duration of insects lifespan. These observations suggest that genome of the nerve cells appears to be a probable initial substrate of Drosophila aging.     

Refractory giant cell arteritis with spinal cord infarction

We report an elderly patient with aggressive steroid-refractory giant cell arteritis manifesting as myelopathy and bilateral visual loss while on treatment. Pathologically, spinal cord infarction was observed and was due to extensive necrotizing granulomatous arteritis of spinal arteries. Spinal cord damage in giant cell arteritis is rare. One prior autopsy report of spinal cord infarction in giant cell arteritis did not identify vasculitic changes in the spinal arteries.     

MHCPEP, a database of MHC-binding peptides: update 1996

MHCPEP is a curated database comprising over 9000 peptide sequences known to bind MHC molecules. Entries are compiled from published reports as well as from direct submissions of experimental data. Each entry contains the peptide sequence, its MHC specificity and, when available, experimental method, observed activity, binding affinity, source protein, anchor positions and publication references. The present format of the database allows text string matching searches but can easily be converted for use in conjunction with sequence analysis packages. The database can be accessed via Internet using WWW, FTP or Gopher.     

Allelopathic effects ofPinus densiflora on undergrowth of red pine forest

Correlation between the distributional frequency of undergrowth species of red pine forest and their germination and growth effected by pine extracts and leachates was found. It was made clear by germination and growth tests that pine toxic substances inhibit the germination and growth of low frequency species more than high frequency species in a red pine forest and that these substances are contained in descending concentration in fresh and fallen leaves, roots, pine forest soil, and pine rain. The concentration of pine toxic substances in extracts or leachates was affected by extracting or leaching within a given period of time, requiring a few hours for extracts or a few days for leachates. The amount of dry weight inhibition of the undergrowth treated by pine leachates was expressed as a growth inhibition index (GII) for the comparison of tolerance in various species. GII is a relative value (%) of the test groups against the control and it is an exponential function of the amount of pine toxic substances affecting the dry weight of the undergrowth. The substances were analyzed by paper and gas chromatography. Benzoic acid and 11 phenolic acids were identifed by gas chromatography. Benzoic acid was considered to be a key factor of allelopathy in the red pine forest.     

Overexpression of leukotriene C4 synthase in bronchial biopsies from patients with aspirin-intolerant asthma

Aspirin causes bronchoconstriction in aspirin-intolerant asthma (AIA) patients by triggering cysteinyl-leukotriene (cys-LT) production, probably by removing PGE2-dependent inhibition. To investigate why aspirin does not cause bronchoconstriction in all individuals, we immunostained enzymes of the leukotriene and prostanoid pathways in bronchial biopsies from AIA patients, aspirin-tolerant asthma (ATA) patients, and normal (N) subjects. Counts of cells expressing the terminal enzyme for cys-LT synthesis, LTC4 synthase, were fivefold higher in AIA biopsies (11.5+/-2.2 cells/mm2, n = 10) than in ATA biopsies (2.2+/-0.7, n = 10; P = 0. 0006) and 18-fold higher than in N biopsies (0.6+/-0.4, n = 9; P = 0. 0002). Immunostaining for 5-lipoxygenase, its activating protein (FLAP), LTA4 hydrolase, cyclooxygenase (COX)-1, and COX-2 did not differ. Enhanced baseline cys-LT levels in bronchoalveolar lavage (BAL) fluid of AIA patients correlated uniquely with bronchial counts of LTC4 synthase+ cells (rho = 0.83, P = 0.01). Lysine-aspirin challenge released additional cys-LTs into BAL fluid in AIA patients (200+/-120 pg/ml, n = 8) but not in ATA patients (0. 7+/-5.1, n = 5; P = 0.007). Bronchial responsiveness to lysine-aspirin correlated exclusively with LTC4 synthase+ cell counts (rho = -0.63, P = 0.049, n = 10). Aspirin may remove PGE2-dependent suppression in all subjects, but only in AIA patients does increased bronchial expression of LTC4 synthase allow marked overproduction of cys-LTs leading to bronchoconstriction.