PurposePrimary congenital glaucoma (PCG), a severe form of glaucoma that presents early in life, is an autosomal recessive eye disorder that results from defects in anterior eye segment. Null mutations in LTBP2 were reported in patients with PCG in Pakistani and Iranian families. This study was aimed to identify the mutation profile of the LTBP2 gene in north Indian patients with PCG.MethodsAfter ethical clearance, 54 unrelated patients with PCG who were either negative or heterozygous for MYOC, CYP1B1, and FOXC1 mutations and 50 ethnically matched non-glaucomatous controls were recruited for the study. PCG diagnosis was established by the presence of buphthalmos in at least one affected eye and associated high intraocular pressure before the age of 3 years. LTBP2 was screened in genomic blood DNA for mutations, with PCR and direct sequencing of PCR amplified fragments.ResultsWe observed one intronic single nucleotide polymorphism (rs3742793) between exons 6 and 7 in the LTBP2 gene in 18 patients with PCG. This nucleotide change resulted in cytosine (C) being replaced by guanosine (G) at position g.75070493. No pathogenic variants were identified in the LTBP2 gene in our cohort of patients.ConclusionsLTBP2 gene mutations are not involved in the pathogenesis of primary congenital glaucoma in our patients. Thus, it is important to screen other glaucoma-associated loci and genes for involvement in congenital glaucoma in cases that are either negative or heterozygous for MYOC, CYP1B1, and FOXC1 mutations to have better insight into the disease pathogenesis.

Deoiled rapeseed lecithin was fractionated with ethanol, and optimum conditions have been determined to improve purified lecithin yield and phosphatidylcholine (PC) enrichment. The effect of extraction time, solvent volume, ethanol concentration and temperature on the yield and the PC enrichment have been described in the form of regression equations. A full factorial experiment method and a second~rder orthogonal design were used in the study. The regression equations were calculated for the maximum value of the response functions optimized by an electronic data processing method, and the results (yield and PC enrichment calculated from regression equations) were compared with those obtained in control experiments. The use of calculated optimal parameters in the fractionation process led to 81-96% and 58% increments in yield and PC enrichment, respectively. KEY WORDS: Alcohols, extreme experiments design, lecithin, lecithin fractionation, optimization of extraction process, phosphatidylcholine, rapeseed lecithin. Upon the basis of apparent changes in extraction yields and PC contents with temperature (Part D, Fig. 1), as well

Optimization by full factorial design of the emulsifying properties of ethanol insoluble fraction from rapeseed lecithinRapeseed lecithin ethanol insoluble fraction (LEIF), was acetylated and optimum conditions for improving LEIF emulsifying property were determined. The effects of reaction time, pyridine amount and temperature on phosphatidyl ethanolamine conversion degree (PECD) and emulsion stability index (ESI) have been described in the form of regression equations. A full factorial design method was used. Regression equations for the maximum value of response functions and computation results were calculated and compared with those obtained in control experiments. Acetylation of rapeseed LEIF with the use of calculated optimal parameters led to 35% and 60% increments in PECD and ESI, respectively.Keywords: Rapeseed lecithin, full factorial design, optimizing of lecithin functionality, N-acetylphosphatidyl ethanolamine, lecithin emulsifying properties.Correspondence: Marian Sosada, Chair and Department of Pharmaceutical Technology, Silesian Medical University, ul. Jagiellońska 4, 41-200 Sosnowiec, Poland. Phone: +48-32-2925548, Fax: +48-32-2667860; e-mail: msosada@slam. katowice.pl were obtained from Sigma (St. Louis, MO, USA). All other solvents and reagents were of analytical grade. Phospholipid acetylationTheoretically, when lecithin acetylation is carried out in the presence of tertiary amine, approximately one mole of acetic anhydride per mole of free amino groups in lecithin is adequate. However, for acetylation in the absence of tertiary amine, a larger quantity is required. Acetylation of plant lecithin is generally carried out with 2-5% addition of acetic anhydride. The amount of acetylating agent depends on PE content of the starting material [3,9,13]. Taking the PE content in rapeseed LEIF into consideration, 50 mmol of acetic anhydride per 100 g LEIF (1.47 mol acetic anhydride per mol PE) was used in acetylation experiments. This amount was maintained in all experiments.A sample of rapeseed LEIF (10 g) and acetone (30 ml) were introduced into a flask fitted with a stirrer and reflux condenser, and placed in an ultrathermostat. Acetic anhydride (50 mmol) and an appropriate amount of pyridine (10-30 mmol/100 g) were added at temperatures between 10 to 40 °C in individual trials (Tab. 1 and 2). Each run of LEIF acetylation was carried out for the set of parameters described in the experimental matrix (Tab. 2). In this matrix, values +1 and -1 correspond to high and low level reaction time, temperature and pyridine amounts, respectively. After the reaction was complete (time 10-50 min, Tab. 2), the final mixture was immediately cooled and acetone, excess acetic anhydride and pyridine, and acetic acid generated during acetylation were quickly removed by distillation under reduced pressure (40 °C, 13.3 hPa). The solid mixture was vigorously stirred with acetone (50 ml) and the precipitate of finely divided solid was filtered through a sintered glass funnel under the vacuum of a water pump, washed t...

rapeseed lecithins obtained from raw lecithin and from lecithin wet gum were evaluated. The chemical composition, fatty acids and metals Die chemischen und ptysikalischen Eigenschaften von Rapslecithin unterschiedlichen Qualitatsgrades, das aus rohem Lecithin und hydratisiertem Lecithin erhalten worden war. wurde bewertet, Die chemicontents, acid, peroxide and iodine values were determined. From the data it could be suggested, that the purified rapeseed lecithins obwet gum are the interesting and valuable material for use in Pharmacy, food industry and cosmetics. sche Zusammensetzung, der Gehalt von Fettsauren und Metallen sowie die Saure-, Peroxid-und Iodzahl wurden bestimmt. Aufgrund diethin, das aus 00-Rapssorten und direkt aus hydratisiertem Rapsol erhalten wurde, ein interessantes und wertvolles Produkt zur Verwcndung in der Pharmazie, Nahrungsmittelindustrie und Kosmetik ist. tained from OO-varieties, especially prepared directly from rapeseed ser Kenndaten kann angenommen werden, daB gereinigtes Raps1cci-

The raw commercial rapeseed lecithin free of erucic acid and glucosinolate (00‐type) was purified by deoiling with acetone and extracting with ethanol. The increasing of phosphatidylcholine (PC) content (from 50 to 70–80%) in obtained rapeseed lecithin extract was performed. It was done by column chromatographic fractionation using a low silica gel–lecithin extract ratio about 2:1 and the various temperatures up to 60°C. The fractions were eluted with 95% ethanol. Effect of rapeseed lecithin column fractionation temperatures is significant and the better purification quality in the higher temperatures was observed. Based on performed investigations it could be summarized that the low silica gel–lecithin extract ratio (2:1) allows to obtain rapeseed lecithin with above 80% content of PC and the best results for temperature 60°C were observed.