A case of sarcomatous degeneration of both ovaries. Successful laparotomy with relative recovery

Peasant woman 27 y.o., virgo intacta, in good general health. Menstruation from 15 liters, not abundant, after 28 days, 3-4 days. About six months ago - the appearance of menorrhagia, but during the last 5 months there is no blood at all. 5 months ago, the patient covered up a swelling on the left side of the abdomen, which was rapidly increasing.

DDIS-21. IN VITRO MICRODIALYSIS RECOVERY OF TRAMETINIB

BACKGROUND Dysregulation of the mitogen-activated protein kinase and phosphoinositide 3-kinase (PI3K) pathways is common in several primary brain tumors and metastatic cancers affecting the central nervous system. Trametinib, and oral MEK1/2 inhibitor, has been effective in preclinical studies against a variety of cell lines and improves survival in select patients with gliomas. Unfortunately, limited and often short-lived benefits are frequently seen in patients. A potential explanation for this is insufficient blood brain barrier penetration at therapeutic concentrations. The primary objective of this study is to measure the in vitro relative recovery of trametinib using microdialysis to establish the feasibility of characterizing its intratumoral pharmacokinetics in vivo using this technique. METHODS In vitro recovery experiments were performed utilizing commercially available microdialysis instruments. Two perfusion rates (0.5–1µL/min), the use of iso- and hypertonic perfusates (artificial CSF and 10% bovine serum albumin in PBS), and the addition of dimethyl sulfoxide (DMSO) were evaluated using the extraction efficiency method. Liquid chromatography-tandem mass spectrometry was used to measure the concentrations of trametinib in stock solutions, dialysate samples, and controls over time. RESULTS Dialysate was initially collected from a solution spiked with 15ng/mL at a rate of 1µL/min for 60 minutes, the recovery was 0.3ng/mL (0.2%) in two samples and below the limit of quantification (< 0.2ng/mL) in three. Dialysate samples were then collected from a stock solution with 150ng/mL of trametinib, recovery ranged from 0.3–15.3ng/mL (0.2–10.2%) in nine detectable samples out of sixteen. Recovery was greater with the addition of DMSO to the perfusate and with the use of a hypertonic perfusate. CONCLUSIONS The poor relative recovery of trametinib using microdialysis suggests that this technique would be unreliable in in vivo studies. Trametinib has poor aqueous solubility, limiting the ability of characterizing its pharmacokinetic profile through microdialysis.

Validation of an ampicillin selection protocol to enrich for mutants of Listeria monocytogenes unable to replicate on fresh produce

ABSTRACT Several outbreaks of listeriosis have implicated fresh produce but genetic factors required for growth of Listeria monocytogenes on produce remain poorly characterized. Based on the fact that β-lactam antibiotics only kill bacterial cells that are growing, we hypothesized that ampicillin selection can enrich for L. monocytogenes mutants unable to grow on produce. For validation, we examined relative recovery of L. monocytogenes strain 2011L-2858 and its cold-sensitive mutant L1E4 following inoculation of cantaloupe rind fragments with 1:1 mixture of the strains and incubation at 4°C with or without ampicillin. Listeria monocytogenes from rind fragments inoculated with the mixed cultures and incubated in the presence of ampicillin were used to inoculate fresh rind fragments for a second round of enrichment. In the presence of ampicillin, the proportion of L1E4 increased from 55% on day 0 to 78% on day 14, with higher recovery (85% after 14 days) in the second round of enrichment. These data suggested that L1E4 was enriched on cantaloupe rind fragments while growing cells of the wildtype were killed by ampicillin. Application of this protocol to transposon mutant libraries from three L. monocytogenes strains yielded several mutants unable to grow on cantaloupe. Thus, ampicillin selection can facilitate discovery of genes essential for growth of L. monocytogenes on fresh produce.

Determination of Nicotine Extracted from Eggplant and Green Pepper by HPLC

Nicotine was separated from eggplant and green pepper seeds (Solanaceous) by High Performance Liquid Chromatography (HPLC).The concentration of nicotine in the eggplant extract (0.871-0.877 μg/ml) was determined by injecting standard material with 0.5 and 1.5 μg/ml, while the concentrations of nicotine in green pepper extract (0.613-0.618 μg/ml) was determined when the standard material was injected with 0.5 and 1.5 μg/ml. The qualitative chemical data was calculated from derivations of the standard material. Nicotine concentration was measured qualitatively in both extracts through the calibration curve and method of the standard addition. This technique has high accuracy and compatibility, bringing the proportion of relative recovery percentage of nicotine extracted from eggplant seeds to 95.880-103.104% and relative error of -4.119-3.104%. Relative recovery percentage of nicotine extracted from green pepper seeds was 97.284 -103.518 % and relative error was -2.716-3.518%.