Comparative Lipidomic Study of Human Milk from Different Lactation Stages and Milk Formulas.

In this report, we present a detailed comparison of the lipid composition of human milk (HM) and formula milk (FM) targeting different lactation stages and infant age range. We studied HM samples collected from 26 Polish mothers from colostrum to 19 months of lactation, along with FM from seven brands available on the Polish market (infant formula, follow-on formula and growing-up formula). Lipid extracts were analysed using liquid chromatography coupled to high-resolution mass spectrometry (LC-Q-TOF-MS). We found that the lipid composition of FM deviates significantly from the HM lipid profile in terms of qualitative and quantitative differences. FM had contrasting lipid profiles mostly across brands and accordingly to the type of fat added but not specific to the target age range. The individual differences were dominant in HM; however, differences according to the lactation stage were also observed, especially between colostrum and HM collected in other lactation stages. Biologically and nutritionally important lipids, such as long-chain polyunsaturated fatty acids (LC-PUFAs) containing lipid species, sphingomyelines or ether analogues of glycerophosphoethanoloamines were detected in HM collected in all studied lactation stages. The observed differences concerned all the major HM lipid classes and highlight the importance of the detailed compositional studies of both HM and FM.

Dietary pomegranate peel improves milk quality of lactating ewes: Emphasis on milk fat globule membrane properties and antioxidative traits.

Concentrated pomegranate peel extract (CPE) was supplemented to ewes, and milk yield and fat content-fatty acid (FA) and phospholipid (PL) composition-were monitored. CPE-fed ewes had higher milk yield, and fat, protein and lactose contents than controls. Milk PL content-20% higher in the CPE-supplemented group-was regulated by treatment and not by total fat content; milk phosphatidylethanolamine and phosphatidylcholine increased by 22 and 26%, respectively, in CPE-supplemented vs. control ewes. Milk saturated FA concentration was higher, and total polyunsaturated and monounsaturated FA content lower in the CPE vs. control group, regardless of milk total fat content. CPE supplementation increased milk antioxidant capacity, suggesting antioxidant transfer from dietary source to milk, increasing stability and nutritive value. Our study provides first evidence for milk quality improvement in terms of antioxidants and PL enrichment without compromising total milk fat, suggesting strategies to improve dairy animals' milk composition without compromising total production.

Dietary oils modify lipid molecules and nutritional value of fillet in Nile tilapia: A deep lipidomics analysis.

The nutritional value of fish fillet can be largely affected by dietary oils. However, little is known about how dietary oils modify lipid molecules in fish fillets. Through biochemical and lipidomics assays, this study demonstrated the molecular characteristics of fillet lipids in Nile tilapia fed with different oils for six weeks. High 18:2n-6 and low 18:3n-3 deposition in phosphoglycerides resulted high 18:2n-6/18:3n-3 ratio in tilapia. Dietary n-3 VLCUFAs intake increased its deposition at sn-1/3 of triglycerides and at sn-2 of phosphatidylcholines. Irrespective of dietary oil, 16:0 was distributed preferentially at the outer positions of glycerol backbone. High 18:2n-6 accumulated at sn-2 position for fish fed with n-3 PUFA-enriched oils. High 18:3n-3 deposited at sn-1/3 in TG, sn-1 in phosphatidylethanolamines, while at sn-2 in phosphatidylcholines. Together, dietary oils change the composition and positional distribution of fatty acids on the glycerol backbone, and change nutritional value of fish for human health.

Terrabacter ginsengisoli sp. nov., isolated from ginseng cultivating soil.

A Gram-positive, strictly aerobic, nonmotile, yellowish, coccus-rod-shaped bacterium (designated Gsoil 653T) isolated from ginseng cultivating soil was characterized using a polyphasic approach to clarify its taxonomic position. The strain Gsoil 653T exhibited optimal growth at pH 7.0 on R2A agar medium at 30°C. Phylogenetic analysis based on 16S rRNA gene sequence similarities, indicated that Gsoil 653T belongs to the genus Terrabacter of the family Humibacillus, and was closely related to Terrabacter tumescens DSM 20308T (98.9%), Terrabacter carboxydivorans PY2T (98.9%), Terrabacter terrigena ON10T (98.8%), Terrabacter terrae PPLBT (98.6%), and Terrabacter lapilli LR-26T (98.6%). The DNA G + C content was 70.5 mol%. The major quinone was MK-8(H4). The primary polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidyl-ethanolamine. The predominant fatty acids were iso-C15:0, iso-C16:0, iso-C14:0, and anteiso-C15:0, as in the case of genus Terrabacter, thereby supporting the categorization of strain Gsoil 653T. However, the DNA-DNA relatedness between Gsoil 653T and closely related strains of Terrabacter species was low at less than 31%. Moreover, strain Gsoil 653T could be both genotypically and phenotypically distinguished from the recognized species of the genus Terrabacter. This isolate, therefore, represents a novel species, for which the name Terrabacter ginsengisoli sp. nov. is proposed with the type strain Gsoil 653T (= KACC 19444T = LMG 30325T).

Flavobacterium panacisoli sp. nov., isolated from soil of a ginseng field.

A novel bacterial strain, designated DCY70(T), was isolated from soil of a ginseng field in Republic of Korea and was characterized in order to determine its taxonomic position. The strain was Gram-reaction negative, yellow-pigmented, rod-shaped and catalase- and oxidase-positive. Based on the 16S rRNA gene sequence similarity, strain DCY70(T) was shown to belong to the genus Flavobacterium, most closely related to Flavobacterium oncorhynchi 631-08(T) (98.4 %), Flavobacterium plurextorum 1126-1H-08(T) (97.9 %), Flavobacterium chilense LM-09-Fp(T) (97.9 %) and Flavobacterium chungangense CJ(T) (97.7 %). The chemotaxonomic characteristics showed only menaquinone-6 (MK-6), iso-C15:0, iso-C15:0 3OH, iso-C17:0 3OH and summed feature 3 as major cellular fatty acids. Polar lipids were phosphatidylethanolamine (PE), two unidentified aminolipids, four unidentified polar lipids and one unidentified phospholipid. The DNA G+C content was 34.9 mol%. Based on the phylogenetic, phenotypic and genotypic data, a novel species, Flavobacterium panacisoli sp. nov., is proposed (=KCTC 32393(T) = JCM 19162(T)).

Identification and quantification of aminophospholipid molecular species on the surface of apoptotic and activated cells.

This protocol measures externalization of aminophospholipids (APLs) to the outside of the plasma membrane using mass spectrometry (MS). APL externalization occurs in numerous events, and it is relevant for transplant medicine, immunity and cancer. In this protocol, externalized APLs are chemically modified by using a cell-impermeable reagent (sulfo-NHS-biotin), and then they are isolated via a liquid:liquid extraction and quantified by reverse-phase liquid chromatography tandem MS (LC-MS/MS) against in-house-generated standards. This protocol describes a complementary method to existing assays that are not quantitative (e.g., annexin V flow cytometry), and it is applicable to the study of membrane reorganization in all cell types during apoptosis (e.g., during development, cancer, psychiatric disorders and other conditions, aging, vesiculation and cell division). The protocol takes ∼2-4 d, including the generation of standards.

Sediminibacterium ginsengisoli sp. nov., isolated from soil of a ginseng field, and emended descriptions of the genus Sediminibacterium and of Sediminibacterium salmoneum.

A Gram-negative bacterium, designated DCY13(T), was isolated from soil of a ginseng field in South Korea. Comparative analysis of 16S rRNA gene sequences showed that strain DCY13(T) shared the highest sequence similarity (95.0 %) with Sediminibacterium salmoneum NBRC 103935(T) and 87.6-91.4 % sequence similarity with other members of the family Chitinophagaceae. Cells were non-spore-forming rods, catalase- and oxidase-positive, motile by gliding and facultatively anaerobic. The only respiratory quinone was menaquinone 7 (MK-7) and the major fatty acids were iso-C17 : 0 3-OH, iso-C15 : 0 and iso-C15 : 1 G. The G+C content of the genomic DNA was 47.5±1.0 mol%. In addition to phosphatidylethanolamine, the major polar lipids were two unidentified aminophospholipids, one unidentified aminolipid and three unidentified polar lipids. The major cell-wall sugars were ribose, xylose and galactose. It is proposed that strain DCY13(T) represents a novel species in the genus Sediminibacterium, for which the name Sediminibacterium ginsengisoli sp. nov. is proposed. The type strain is DCY13(T) ( = KCTC 12833(T)  = JCM 15794(T)  = DSM 22335(T)). Emended descriptions of the genus Sediminibacterium and of Sediminibacterium salmoneum are also proposed.

Pedobacter luteus sp. nov., isolated from soil.

Two strains of Gram-staining-negative, rod-shaped bacteria that were motile by gliding, N7d-4(T) and B4a-b5, were isolated during a study of culturable bacteria in soil cultivated with potatoes. These isolates grew at 15-37 °C and at pH 6.5-7.0. The major cellular fatty acids were summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1ω7c), anteiso-C15 : 0, iso-C15 : 0, iso-C17 : 0 3-OH and iso-C17 : 1ω9c. The major polar lipids were phosphatidyl-N-methylethanolamine and phosphatidylethanolamine. The strains contained d-18 : 0 and d-19 : 0 sphingosines. The DNA G+C contents of strains N7d-4(T) and B4a-b5 were 48.5 and 46.9 mol% (HPLC), respectively. A phylogenetic analysis based on 16S rRNA gene sequences showed that strains N7d-4(T) and B4a-b5 were affiliated with Pedobacter species in the family Sphingobacteriaceae. Strains N7d-4(T) and B4a-b5 shared 99.9 % sequence similarity, and the most closely related Pedobacter type strains were Pedobacter composti TR6-06(T) (96.5 and 96.7 % sequence similarity, respectively), P. oryzae N7(T) (95.4 and 95.6 %) and P. caeni LMG 22862(T) (94.0 and 94.4 %). Phenotypic data and phylogenetic inference clearly distinguished the two isolates from other Pedobacter species. Based on these data, the isolates are considered to represent a novel species of the genus Pedobacter, for which the name Pedobacter luteus sp. nov. is proposed. The type strain is N7d-4(T) ( = KCTC 22699(T)  = DSM 22385(T)).

Pedobacter ginsengiterrae sp. nov., isolated from soil of a ginseng field.

A Gram-stain-negative, oxidase- and catalase-positive bacterial strain that was motile by gliding and produced a pink pigment, designated DCY49(T), was isolated from soil of a ginseng field in a mountainous region of Chungbuk province, South Korea. 16S rRNA gene sequence analysis revealed that strain DCY49(T) belonged to the genus Pedobacter (93.0-96.3 % similarity). Strain DCY49(T) contained MK-7 as the predominant menaquinone. The major fatty acids were summed feature 3 (containing C16 : 1ω7c, C16 : 1ω6c and/or iso-C15 : 0 2-OH), iso-C15 : 0, iso-C17 : 0 3-OH and C16 : 0, and the main polar lipid was phosphatidylethanolamine. The G+C content of the genomic DNA of strain DCY49(T) was 40.5 mol%. Strain DCY49(T) differed from related Pedobacter species by a number of phenotypic characteristics. On the basis of data from the present polyphasic study, strain DCY49(T) is described as representing a novel species of the genus Pedobacter, for which the name Pedobacter ginsengiterrae sp. nov. is proposed. The type strain is DCY49(T) ( = KCTC 23317(T) = JCM 17338(T)).

Poxvirus membrane biogenesis: rupture not disruption.

Enveloped viruses acquire their membrane from the host by budding at, or wrapping by, cellular membranes. Transmission electron microscopy (TEM) images, however, suggested that the prototype member of the poxviridae, vaccinia virus (VACV), may create its membrane 'de novo' with free open ends exposed in the cytosol. Within the frame of the German-wide priority programme we re-addressed the biogenesis and origin of the VACV membrane using electron tomography (ET), cryo-EM and lipid analysis of purified VACV using mass spectrometry (MS). This review discussed how our data led to a model of unconventional membrane biogenesis involving membrane rupture and the generation of a single open membrane from open membrane intermediates. Lipid analyses of purified virus by MS suggest an ER origin with a relatively low cholesterol content compared with whole cells, confirming published data. Unlike previous reports using thin-layer chromatography, no depletion of phosphatidylethanolamine was detected. We did detect, however, an enrichment for phosphatidic acid, diacylglycerol and phosphatidylinositol in the virion. Our data are discussed in the light of other pathogens that may requirecellular membrane rupture during their intracellular life cycle.

The use of a new, modified Dittmer-Lester spray reagent for phospholipid determination by the TLC image analysis technique.

A new phospholipid-specific spray reagent is described. A new phospholipid-specific spray reagent, which is a modification of the Dittmer-Lester reagent, is described in authors' studies. The difference between these two reagents is in the addition of tin (II) chloride to the proposed spray reagent. The use of the described spray reagent together with an image analysis technique allows not only for qualitative, but also for quantitative, determination of major phospholipid classes. Separation of phosphatidylserine (PS), phosphatidylethanolamine (PE) and phosphatidylcholine (PC) was conducted on an HPTLC plate with a mixture of chloroform, methanol and 25% ammonia solution in a volume ratio of 65:25:4 as mobile phase. The calibration curves were linear in the ranges of 5.0-25.0, 1.5-15.0 and 1.0-20.0 µg/spot for PC, PS and PE, respectively. The use of the new spray reagent resulted also in lower limits of detection than the standard molybdenum method for the investigated phospholipids. The proposed method was used to determine the amount of PS in the dietary supplement 'Session', and of PS, PE and PC in biological samples, with good results.

Characterization of lecithin isolated from anchovy (Engraulis japonica) residues deoiled by supercritical carbon dioxide and organic solvent extraction.

UNLABELLED: Lecithin was isolated and characterized from anchovy (Engraulis japonica) deoiled residues using supercritical carbon dioxide (SC-CO(2)) at a semibatch flow extraction process and an organic solvent (hexane) extraction. SC-CO(2) extraction was carried out to extract oil from anchovy at different temperatures (35 to 45 °C) and pressures (15 to 25 MPa). Extraction yield of oil was influenced by physical properties of SC-CO(2) with temperature and pressure changes. The major phospholipids of anchovy lecithin were quantitatively analyzed by high-performance liquid chromatography. Phosphatidylcholine (PC) (68%± 1.00%) and phosphatidylethanolamine (PE) (29%± 0.50%) were the main phospholipids. Thin layer chromatography was performed to purify the individual phospholipids. The fatty acid compositions of lecithin, PC, and PE were analyzed by gas chromatography. A significant amount of eicosapentaenoic acid and docosahexaenoic acid were present in both phospholipids of PC and PE. Emulsions of lecithin in water were prepared through the use of a homogenizer. Oxidative stability of anchovy lecithin was high in spite of its high concentration of long-chain polyunsaturated fatty acids. PRACTICAL APPLICATION: Lecithin can be totally metabolized by humans, so is well tolerated by humans and nontoxic when ingested. Lecithin from anchovy contain higher amounts of ω-3 fatty acids especially EPA and DHA, it may have positive outcome to use in food and pharmaceutical industries.

Determination of phosphatidylethanolamine molecular species in various food matrices by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS2).

A liquid chromatographic-electrospray ionization-tandem mass spectrometric (LC-ESI-MS(2)) method has been developed for determination of the molecular species of phosphatidylethanolamine (PE) in four food matrices (soy, egg yolk, ox liver, and krill oil). The extraction and purification method consisted of a pressurized liquid extraction procedure for total lipid (TL) extraction, purification of phospholipids (PLs) by adsorption on a silica gel column, and separation of PL classes by semi-preparative normal-phase HPLC. Separation and identification of PE molecular species were performed by reversed-phase HPLC coupled with electrospray ionization tandem mass spectrometry (ESI-MS(2)). Methanol containing 5 mmol L(-1) ammonium formate was used as the mobile phase. A variety of PE molecular species were detected in the four food matrices. (C16:0-C18:2)PE, (C18:2-C18:2)PE, and (C16:0-C18:1)PE were the major PE molecular species in soy. Egg yolk PE contained (C16:0-C18:1)PE, (C18:0-C18:1)PE, (C18:0-C18:2)PE, and (C16:0-C18:2)PE as the major molecular species. Ox liver PE was rich in the species (C18:0-C18:1)PE, (C18:0-C20:4)PE, and (C18:0-C18:2)PE. Finally, krill oil which was particularly rich in (C16:0(alkyl)-C22:6(acyl))plasmanylethanolamine (PakE), (C16:0-C22:6)PE, and (C16:0-C20:5)PE, seemed to be an interesting potential source for supplementation of food with eicosapentaenoic acid and docosahexaenoic acid.

Inverse association between trans isomeric and long-chain polyunsaturated fatty acids in pregnant women and their newborns: data from three European countries.

BACKGROUND: trans unsaturated fatty acids are thought to interfere with essential fatty acid metabolism. To extend our knowledge of this phenomenon, we investigated the relationship between trans isomeric and long-chain polyunsaturated fatty acids (LCPUFA) in mothers during pregnancy and in their infants at birth. METHODS: Fatty acid composition of erythrocyte phosphatidylcholine (PC) and phosphatidylethanolamine (PE) was determined in Spanish (n = 120), German (n = 78) and Hungarian (n = 43) women at the 20th and 30th week of gestation, at delivery and in their newborns. RESULTS: At the 20th week of gestation, the sum of trans fatty acids in PE was significantly (p < 0.01) lower in Hungarian [0.73 (0.51), % wt/wt, median (IQR)] than in Spanish [1.42 (1.36)] and German [1.30 (1.21)] women. Docosahexaenoic acid (DHA) values in PE were significantly (p < 0.01) higher in Hungarian [5.65 (2.09)] than in Spanish [4.37 (2.60)] or German [4.39 (3.3.2)] women. The sum of trans fatty acids significantly inversely correlated to DHA in PCs in Spanish (r = -0.37, p < 0.001), German (n = -0.77, p < 0.001) and Hungarian (r = -0.35, p < 0.05) women, and in PEs in Spanish (r = -0.67, p < 0.001) and German (r = -0.71, p < 0.001), but not in Hungarian (r = -0.02) women. Significant inverse correlations were seen between trans fatty acids and DHA in PEs at the 30th week of gestation (n = 241, r = -0.52, p < 0.001), at delivery (n = 241, r = -0.40, p < 0.001) and in cord lipids (n = 218, r = -0.28, p < 0.001). CONCLUSION: Because humans cannot synthesize trans isomeric fatty acids, the data obtained in the present study support the concept that high maternal trans isomeric fatty acid intake may interfere with the availability of LCPUFA both for the mother and the fetus.

Isolation and characterization of lecithin from squid (Todarodes pacificus) viscera deoiled by supercritical carbon dioxide extraction.

UNLABELLED: Marine lecithin was isolated and characterized from squid (Todarodes pacificus) viscera residues deoiled by supercritical carbon dioxide (SC-CO(2)) extraction. SC-CO(2) extraction was carried out to extract the oil from squid viscera at different temperatures (35 to 45 °C) and pressures (15 to 25 MPa). The extraction yield was higher at highest temperature and pressure. The major phospholipids of squid viscera lecithin were quantified by high-performance liquid chromatography (HPLC). Phosphatidylcholine (PC; 80.5% ± 0.7%) and phosphatidylethanolamine (PE; 13.2% ± 0.2%) were the main phospholipids. Thin layer chromatography (TLC) was performed to purify the individual phospholipids. The fatty acid compositions of lecithin, PC and PE were analyzed by gas chromatography (GC). A significant amount of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were present in both phospholipids of PC and PE. Emulsions of lecithin in water were prepared through the use of a homogenizer. The oxidative stability of squid viscera lecithin was high in spite of its high concentration of long-chain polyunsaturated fatty acids. PRACTICAL APPLICATION: Squid viscera are discarded as a waste by fish processing industry. Since lecithin from squid viscera contains higher amounts of polyunsaturated fatty acids, it may have promising effect to use in food, pharmaceutical, and cosmetic industries.

Testes of obese rats are highly responsive to n-3 long-chain fatty acids.

The present study investigated whether fatty acid compositions of testes are affected by the obese condition and dietary n-3 long-chain fatty acid (LCFA) intake. Male lean and obese Zucker rats were fed a 15 % (w/w, total diet) fat diet containing either 0 or 5·0 % (w/w, total fatty acids) n-3 LCFA for 8 weeks. Reproductive organ weights, sperm morphology and fatty acid composition of phosphatidylcholine (PC), and phosphatidylethanolamine (PE) of testes were analysed. The obese rats had significantly (P < 0·0001) smaller epididymides and seminal vesicles, larger prostates and abundant underdeveloped testes compared with lean rats. Diet treatment did not affect the sex organ weights. The effect of genotype on fatty acid composition was minor in PC and PE except for DHA (22 : 6n-3). The n-3 LCFA diet significantly (P < 0·0001) elevated 22 : 6n-3 and reduced arachidonic acid (20 : 4n-6) and DPA (22 : 5n-6) in testicular PC and PE of lean and obese rats compared with the control diet. The acylation of dietary n-3 LCFA into 22 : 6n-3 was 2-fold higher in obese rat testes than in lean rats fed the same diet. Underdeveloped testes had 70 % less 22 : 5n-6 in PC and PE than normal-size testes. Results indicate that testicular fatty acid composition is sensitive to dietary fat modulations and especially obese rats responded more to dietary n-3 LCFA than their lean counterparts. The selective reduction in 22 : 5n-6 in underdeveloped testes indicates that 22 : 5n-6 is important in male reproduction in rats and requires further study to define the role of elongation and desaturation in testicular development.

Effects of phosphatidylcholine and phosphatidylethanolamine on the photooxidation of canola oil.

Effects of chlorophyll b, phosphatidylcholine (PC), and phosphatidylethanolamine (PE) on the oxidation of canola oil under 1700 lux light at 10 degrees C were studied by determining the headspace oxygen depletion and peroxide value. Chlorophyll b increased the oil oxidation under light via production of singlet oxygen. PC and PE did not affect the headspace oxygen depletion and peroxide formation in the oil in the absence of chlorophyll b at 10 degrees C; however, they increased in the presence of chlorophyll, with higher effect of PC than PE. Chlorophyll was degraded during the oil photooxidation and PC and PE decreased its photodecomposition with higher protection by PC than PE. There was no effect of light or singlet oxygen on PC and PE during the oil photooxidation. The results clearly showed that PC and PE were prooxidants in the photooxidation of canola oil containing chlorophyll b by protecting it from photodecomposition, and thus PC and PE should be removed from the oil containing chlorophyll to improve the photooxidative stability.

The effect of maternal T1DM on the fatty acid composition of erythrocyte phosphatidylcholine and phosphatidylethanolamine in infants during early life.

BACKGROUND: The risk for type 1 diabetes (T1DM) in children of mothers with T1DM is different to that in children of fathers with T1DM. Fatty acid (FA) intake, in particular EPA and DHA, has been associated with T1DM risk and has been suggested to be inadequate in infants of diabetic mothers. We asked, therefore, whether EPA and DHA FA nutritional status in offspring of mothers with T1DM could contribute to their reduced T1DM risk. METHODS: BABYDIET follows children with increased genetic and familial risk for T1DM from birth to age 3 years. FA nutritional state was assessed by determining the erythrocyte membrane phosphatidylethanolamine (PE) and phosphatidylcholine (PC) composition in children of T1DM mothers and children of T1DM fathers or with T1DM siblings participating in the BABYDIET study. Samples for determination of erythrocyte membrane FA composition were collected at ages 3 and 12 months in 48 and 49 infants, respectively. FA measurements were adjusted for breastfeeding duration, FA supplementation, and gluten exposure. RESULTS: 3-months-old children of T1DM mothers and T1DM fathers/sibs had similar levels of PC DHA and EPA (DHA 1.53+/-0.23 vs. 1.65+/-0.11 wt.%; EPA 0.15+/-0.02 vs. 0.21+/-0.03 wt.%) and PE DHA and EPA (DHA 7.54+/-0.37 vs. 7.92+/-0.38 wt.%; EPA 0.53+/-0.06 vs. 0.61+/-0.04 wt.%). No differences were also observed after stratification for breastfeeding. At age 12 months, a minor reduction of PE DHA was observed in children of T1DM mothers. Expected higher levels for DHA and EPA in fully breastfed children and in children of mothers taking fish oil supplementation were observed at 3 months in all children. Other differences included increased levels of the major saturated FA 16:0 in 3-months-old infants from T1DM mothers (PC 35.45+/-0.35 vs. 33.89+/-0.26 wt.%, mean +/- SEM, P(corr)=0.005; PE 16.13+/-0.39 vs. 14.93+/-0.24 wt.%, P(corr)=0.05). CONCLUSION: Although FA status was not identical in children from T1DM mothers and from T1DM fathers, maternal T1DM was not associated with changes in offspring's EPA and DHA incorporation into erythrocyte membrane.

Essential n-3 fatty acids in pregnant women and early visual acuity maturation in term infants.

BACKGROUND: Docosahexaenoic acid (DHA) is important to neural development. Whether DHA intakes are low enough in some pregnant women to impair infant development is uncertain. OBJECTIVE: We sought to determine whether DHA deficiency occurs in pregnant women and contributes to poor infant development. DESIGN: Biochemical cutoffs, dietary intakes, or developmental scores indicative of DHA deficiency are not defined. Infant development has a distribution in which an individual's potential development is unknown. This was a randomized intervention to establish a distribution of developmental scores for infants of women with DHA intakes considered to be above requirements against which to compare the development of infants of mothers consuming their usual diet. DHA (400 mg/d; n = 67) or a placebo (n = 68) was consumed by the women from 16 wk gestation until delivery. We determined maternal red blood cell ethanolamine phosphoglyceride fatty acids, dietary intakes at 16 and 36 wk gestation, and infant visual acuity at 60 d of age. RESULTS: We described an approach to identify DHA deficiency when biochemical and functional markers of deficiency are unknown. In multivariate analyses, infant visual acuity was related to sex (beta = 0.660, SE = 0.93, and odds ratio = 1.93) and maternal DHA intervention (beta = 1.215, SE = 1.64, and odds ratio = 3.37). More infant girls in the placebo than in the DHA intervention group had a visual acuity below average (P = 0.048). Maternal red blood cell ethanolamine phosphoglyceride docosatetraenoic acid was inversely related to visual acuity in boys (rho = -0.37, P < 0.05) and girls (rho = -0.48, P < 0.01). CONCLUSIONS: These studies suggest that some pregnant women in our study population were DHA-deficient.

A high-throughput method for the quantitative analysis of indole-3-acetic acid and other auxins from plant tissue.

To investigate novel pathways involved in auxin biosynthesis, transport, metabolism, and response, we have developed a high-throughput screen for indole-3-acetic acid (IAA) levels. Historically, the quantitative analysis of IAA has been a cumbersome and time-consuming process that does not lend itself to the screening of large numbers of samples. The method described here can be performed with or without an automated liquid handler and involves purification solely by solid-phase extraction in a 96-well format, allowing the analysis of up to 96 samples per day. In preparation for quantitative analysis by selected ion monitoring-gas chromatography-mass spectrometry, the carboxylic acid moiety of IAA is derivatized by methylation. The derivatization of the IAA described here was also done in a 96-well format in which up to 96 samples can be methylated at once, minimizing the handling of the toxic reagent, diazomethane. To this end, we have designed a custom diazomethane generator that can safely withstand high flow and accommodate larger volumes. The method for IAA analysis is robust and accurate over a range of plant tissue weights and can be used to screen for and quantify other indolic auxins and compounds including indole-3-butyric acid, 4-chloro-indole-3-acetic acid, and indole-3-propionic acid.