High-performance liquid chromatography with on-line coupled UV, mass spectrometric and biochemical detection for identification of acetylcholinesterase inhibitors from natural products.

A high-performance liquid chromatography (HPLC) method with on-line coupled ultraviolet (UV), mass spectrometry (MS) and biochemical detection for acetylcholinesterase (AChE) inhibitory activity has been developed. By combining the separation power of HPLC, the high selectivity of biochemical detection, and the ability to provide molecular mass and structural information of MS, AChE inhibitors can be rapidly identified. The biochemical detection was based on a colorimetric method using Ellman's reagent. The detection limit of galanthamine, an AChE inhibitor, in the HPLC-biochemical detection is 0.3 nmol. The three detector lines used, i.e., UV, MS and Vis for the biochemical detection were recorded simultaneously and the delay times of the peaks obtained were found to be consistent. This on-line post-column detection technique can be used for the identification of AChE inhibitors in plant extracts and other complex mixtures such as combinatorial libraries.

Caring for the individual.

People do not lose their individuality as they grow older. With the use of a moving case history Carolyn Best describes how a lifetime's experiences and personality should be taken into account when dealing with older people.

Postoperative nausea and vomiting in paediatric surgical inpatients.

A prospective study of postoperative nausea and vomiting (PONV) was conducted in 415 children presenting for inpatient surgery. The overall incidence of PONV was 18.1%). The highest incidence was in children undergoing ENT procedures and increased with age. Avoidance of intraoperative opioids and the use of local anaesthesia and/or non-steroidal anti-inflammatory drugs reduced the incidence of nausea and vomiting postoperatively.

Hemispheric asymmetries in adults' perception of infant emotional expressions.

Accounts of emotion lateralization propose either overall right hemisphere (RH) advantage or differential RH versus left hemisphere (LH) involvement depending on the negative-positive valence of emotions. Perceptual studies generally show RH specialization. Yet viewer emotional responses may enhance valence effects. Because infant faces elicit heightened emotion in viewers, perceptual asymmetries with chimeric infant faces were assessed. First, it was determined that chimeras must be paired with their counterparts, not their mirror images, to tap viewers' sensitivity to adult facial asymmetries. Results showed an RH perceptual bias for infant cries but bihemispheric sensitivity to asymmetries in infant smiles. This effect was not due to LH featural versus RH holistic processing and held for additional, intensity-matched, spontaneous expressions. Specialized RH sensitivity to infant cries may reflect an evolutionary advantage for rapid response to infant distress.

Modulation of human granulosa cell steroid production in vitro by tumor necrosis factor alpha: implications of white blood cells in culture.

OBJECTIVE: To test the hypothesis that the macrophage cytokine, tumor necrosis factor-alpha (TNF-alpha), directly inhibits progesterone, estrone (E1), and estradiol (E2) synthesis by human granulosa cells in vitro in the presence and absence of white blood cells. METHODS: Granulosa cells from follicle aspirates of patients undergoing in vitro fertilization (IVF) were separated from red blood cells on 50% Percoll columns. Such preparations contained numerous white blood cells (lymphocytes, 40-52%, and macrophages, 6-14%) as determined with immunocytochemistry. In some studies, anti-CD45 magnetic beads followed by an additional adherence step and media change were used to remove white blood cells from granulosa cell cultures. Granulosa cells with and without associated white blood cells were cultured in basal and hCG-supplemented media. Androstenedione (40 ng/mL) and/or recombinant TNF-alpha (0.5-50 ng/mL) were added to triplicate wells. Media were harvested for radioimmunoassay of progesterone, E1, and E2 after 24 and 48 hours of incubation. RESULTS: The effects of TNF-alpha on progesterone production in white blood cell-associated cultures were inconsistent when 0.5 ng/mL TNF-alpha was added under basal conditions. At higher TNF-alpha doses (5-50 ng/mL) and under hCG-stimulated conditions, there was a consistent decrease in progesterone production, but the effect was not clearly dose-dependent. It was possible to remove white blood cells effectively from granulosa cell cultures. In granulosa cell cultures without associated white blood cells, 0.5 ng/mL of TNF-alpha at 48 hours produced an increase in progesterone, whereas 50 ng/mL of TNF-alpha decreased progesterone (P < .001). Estrone and E2 were both decreased by TNF-alpha regardless of whether white blood cells were present in culture, without clear evidence of dose-dependency. Granulosa cell viability and proliferation were unaffected by TNF-alpha as demonstrated by direct cell counts, trypan blue exclusion, and tetrazolium salt viability assays. CONCLUSIONS: In the normal ovary, TNF-alpha may influence the development of the dominant follicle by inhibiting aromatase activity. It may also mediate oocyte maturation disorders and ovarian endocrine dysfunction in some pathologic states. White blood cells can be effectively removed from granulosa cell cultures. Application of this removal technique will facilitate future granulosa cell studies by allowing more precise determination of direct granulosa cell function.

The azaspirane SKF 105685 ameliorates renal allograft rejection in rats.

The azaspirane SKF 105685 (N,N-dimethyl-8, 8-dipropyl-2-azaspiro (4.5) decane-2-propanamine dihydrochloride) has been shown to attenuate or reverse the course of immunologic disease in several animal models, possibly through the induction of nonspecific suppressor activity. To investigate its effects on immune-mediated renal disease, SKF 105685 was administered by gavage to rats with kidney allografts. Six days after transplantation, GFR (inulin clearance, 1.46 +/- 0.27 versus 0.41 +/- 0.15 mL/min per kg; P < 0.005) and RPF (p-aminohippurate clearance, 5.48 +/- 0.98 versus 1.99 +/- 0.72 mL/min per kg; P < 0.01) were significantly higher in SKF 105685-treated rats compared with vehicle-treated control rats. In addition, mononuclear inflammatory cell infiltrates were significantly reduced in SKF 105685-treated animals compared with controls. Treatment also reduced renal production of thromboxane B2 (81 +/- 22 versus 424 +/- 76 pg/min per mg of protein; P < 0.0005), prostaglandin E2 (612 +/- 165 versus 2,059 +/- 351 pg/min per mg of protein; P < 0.005), and 6-keto prostaglandin F1 alpha (217 +/- 56 versus 943 +/- 186 pg/min per mg of protein; P < 0.005), but interleukin-1 beta mRNA levels within kidney allografts were not affected by treatment. Thus, the azaspirane SKF 105685 is a novel immunosuppressive agent that substantially ameliorates renal allograft rejection in the rat. Although the mechanism of action is unknown, the beneficial effects of SKF 105685 in rejection may relate to its ability to induce suppressor activity and/or its effects on eicosanoid production.