Ingestion of ponderosa pine needles by late-pregnant cattle causes premature parturition induced by a profound and progressive decrease in uterine blood flow. In vitro perfused placentomes obtained from late-pregnant cows were used in the bioassay-guided fractionation of Pinus ponderosa pine needles to isolate a new class of vasoactive lipid substances. Chromatographically inseparable mixtures of active compounds were isolated by Soxhlet extraction and flash column chromatography. The structures of intact myristate and/or laurate diesters of 1,14-tetradecanediol or 1,12-dodecanediol in these extracts were determined by lH and 13C NMR, IR, and high-and low-resolution FAB mass spectrometric analyses. Saponification of small samples of isolated diesters, derivatization, and GCiMS verified major components as lauric and myristic acids and 1,14-tetradecanediol. Structures of individual components of mixtures of lipids were confirmed by MS-MS analysis.

Isolation, characterization, and biological evaluation of active components of Colchicum stevenii Kunth (Colchicaceae) are described. Colchicum stevenii is an unexplored Jordanian specie with toxic reputation. Directed by brine shrimp lethality test (BST), methanolic extraction, liquid-liquid partition, preparative TLC, and semi-preparative HPLC, it resulted in the isolation of six cytotoxic compounds. The compounds, reported for the first time from this specie, are: (-)-colchicine (1), 2-demethyl-(-)-colchicine (2), (-)-cornigerine (3), beta-lumicolchicine (4), (-)-isoandrocymbine (5) and (-)-O-methylandrocymbine (6). A new, in-house developed, acidic-based reverse-phase gradient semi-preparative HPLC method for the separation of colchisides is presented here. Structural elucidation was based on spectroscopic techniques principally; 1H-NMR and low resolution EIMS. Based on BST results, reported as LC50 values in microg mL(-1) (ppm) with 95% confidence intervals, (-)-colchicine (2.5 ppm) and (-)-cornigerine (2.7 ppm) were the most potent.

Analysis and distribution of Pb and Cd in different mice organs, including the liver, kidney, spleen, heart, and blood, were evaluated before and after treatment with different aqueous concentrations of Nigella sativa (1.25-10.0 mg/L). Atomic absorption spectrometry was used for analysis of Pb and Cd in these organs. Results indicated that the Pb in the unexposed group of mice without treatment with N. sativa (black cumin) was in the following order: liver > heart > spleen > kidney, and the distribution of Pb in various organs of the unexposed group was not affected significantly by N. sativa. Moreover, results of mice exposed for Pb show that the Pb concentrations in different organs were reduced significantly (p < 0.05) by 72.9%, 63.4%, 72.3%, 66.7%, and 39.5% at a dose of 10 mg/L of N. sativa for the liver, kidney, heart, spleen, and blood, respectively. Furthermore, the distribution of Cd in the unexposed Cd group of mice without treatment with N. sativa was in the following order: kidney > heart > spleen > liver. Nigella sativa at 10 mg/L reduced Cd levels in mice exposed to Cd by 75.5%, 83.3%, 47.0%, 95.3%, and 100% in the liver, kidney, heart, spleen, and blood, respectively, whereas blood Cd concentrations were lowered to below the detection limit of 0.05 mug/L. A 28-d exposure of mice to a Cd-Pb mixture at a concentration of 1 ppm in drinking water induced a highly significant inhibition (p < 0.0001) of antibody response to human serum (80.5%). The suppressed immune responses in mice pretreated with the Cd-Pb mixture were reversed by 43.1% and 38.9% in the presence of 1.25 and 2.5 mg/mL of N. sativa, respectively, whereas higher concentrations (5-10 mg/mL) of N. sativa increased the immunosuppression significantly. Nigella sativa at 1.25-10 mg/mL did not induce any significant modulation of the antibody response in unexposed mice.

Pinus ponderosa needle (PN) ingestion by late pregnant cows results in decreased uterine blood flow, premature parturition, and retained placentae. Further, plasma from PN-fed cows increases caruncular arterial tone (i.e., induces prolonged contraction) in an isolated perfused bovine placentome. A novel class of vasoactive lipids was isolated and identified using a bovine placentome assay-guided fractionation of CH2Cl2 extracts of PN. Placentome perfusion tests indicated that 1-12-dodecanedioyl-dimyristate (14-12-14) was the most potent of the PN lipids for increasing caruncular arterial tone. Late pregnant guinea pigs (GP) were used to evaluate the abortifacient activity of these vasoactive lipids. In Study 1, on d 50 of gestation, part of the control diet was replaced with chopped PN (Diet A) or chopped PN subjected to sequential extraction with diethyl ether (Et2O; Diet B); Et2O and CH2Cl2 (Diet C); and Et2O, CH2Cl2, and methanol (Diet D). The GP on Diets A and B exhibited shorter (P<.01) gestation lengths and reduced (P<.01) pig birth weights than GP on the control diet or Diets C and D. Further, only GP on Diets A and B exhibited retained placentae. In Study 2, on d 50 of gestation, part of the control diet was replaced with chopped PN that had been subjected to exhaustive CH2Cl2 extraction and then infiltrated with either CH2Cl2 alone (Diet E), CH2Cl2 containing 14-12-14 (Diet F), or CH2Cl2 containing isocupressic acid (Diet G); then solvents were evaporated. The GP consuming Diet F had shorter (P<.05) gestation lengths and reduced (P<.05) pig birth weights than did GP consuming Diets E or G. The GP consuming Diet F also exhibited a high incidence of retained placentae. These data provide evidence that a unique class of vasoactive lipids in PN exhibit abortifacient activity in guinea pigs.

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