Anti-inflammatory activity of a lipid fraction (lyprinol) from the NZ green-lipped mussel

Abstract  A lipid-rich extract, preparared by supercritical fluid extraction of fresh stabilized mussel powder (Lyprinol), showed significant anti-inflammatory (AI) activity given therapeutically and prophylactically po to Wistar and Dark Agouti rats developing either (a) adjuvant-induced polyarthritis or (b) collagen(II)-induced autoallergic arthritis, with ED50≤15 mg/kg; c.f. naproxen≥25 mg/kg or various therapeutic oils (flaxseed, evening primrose, fish)≥1800 mg/kg given orally. Lyprinol showed little or no activity in acute irritation assays (carrageenan, kaolin, histamine) indicating it is not mimicking rapid-acting NSAIDs. Incorporating Lyprinol into arthritigenic adjuvants composed of heat-killed Mycobacterium. tuberculosis suspended in olive oil or squalane, effectively prevented arthritis development at a dose of 5 mg/rat. By contrast, ‘dummy adjuvants’ prepared with Mycobacterium tuberculosis and flaxseed, evening primrose or fish oils were still arthritigenic in Dark Agouti rats (doses of oil=90 mg/rat).

Antifeedants in the Feces of the Pine Weevil Hylobius abietis: Identification and Biological Activity

Abstract  Egg-laying females of the pine weevil, Hylobius abietis (L.), regularly deposit feces adjacent to each egg. Egg cavities are gnawed in the bark of roots of recently dead conifer trees. After egg deposition, the cavity is sealed by feces and a plug of bark fragments. Root bark containing egg cavities with feces is avoided as food by pine weevils, which indicates the presence of natural antifeedants. Here we present the first results of the isolation and chemical analyses of antifeedant compounds in the feces of H. abietis. In feeding bioassays, methanol extracts of the feces revealed strong antifeedant properties. Methanol extracts were fractionated by medium-pressure liquid chromatography and the antifeedant effects were mainly found in the fractions of highest polarity. Volatile compounds in the active fractions were identified by gas chromatography–mass spectrometry (GC–MS) and the nonvolatile compounds were characterized by pyrolysis–GC–MS. Based on mass spectra, a number of compounds with various chemical structures were selected to be tested for their antifeedant properties. Antifeedant effects were found among compounds apparently originating from lignin: e.g., a methylanisol, guaiacol, veratrol, dihydroxybenzenes, and dihydroconiferyl alcohol. A weak effect by fatty acid derivatives was found. The types of naturally occurring antifeedant compounds identified in this study may become useful for the protection of planted conifer seedlings against damage by H. abietis.

Antifeedant activity and toxicity of leaf extracts from Porteresia coarctata Takeoka and their effects on the physiology of Spodoptera litura (F.)

Abstract  Antifeeding and toxic effects of chemical extracts from Porteresia coarctata Takeoka on the insect pest Spodoptera litura (F.) were investigated in the laboratory. Hexane extracts of P. coarctata leaves were re-dissolved in dimethyl sulfoxide and applied in water to leaves of Ricinus communis L. (castor). A concentration of 1,000 and 2,000 ppm extract exhibited significant antifeeding and toxic effects to third instar larvae when compared to the same concentration of dimethyl sulfoxide in water as control. One day after feeding on extract-treated castor leaves, the mortality of S. litura larvae was 50%. Mortality increased to 95% after 11 days at a dosage of 2,000 ppm. Compared to the control, third instar larvae fed on castor leaves treated with P. coarctata leaf extract at different concentrations showed significant reduction in protein and DNA content in the fat body and midgut tissues. Results demonstrate the insecticidal and antifeedant properties of P. coarctata leaf extract against the generalist pest S. litura leading to biochemical changes in the insect body. Thus, P. coarctata contains phytochemicals, and the extract, in crude and pure form, has potential as a botanical insecticide in alternative control strategies against Lepidoptera pests.

Antidiabetic effect of novel modulating peptides of G-protein-coupled kinase in experimental models of diabetes

AbstractAims/hypothesis  G-protein-coupled receptor kinases (GRKs) play a key role in agonist-induced desensitisation of G-protein-coupled receptors (GPCRs) that are involved in metabolic regulation and glucose homeostasis. Our aim was to examine whether small peptides derived from the catalytic domain of GRK2 and -3 would ameliorate Type 2 diabetes in three separate animal models of diabetes.

Antidiabetic actions of estrogen: Insight from human and genetic mouse models

Abstract  There is increasing evidence both in humans and rodents linking the endogenous estrogen 17β-estradiol (E2) to the maintenance of glucose homeostasis. Postmenopausal women develop visceral obesity and insulin resistance and are at increased risk for type 2 diabetes mellitus, but hormone replacement therapy leads to a reduction in the incidence of diabetes. In various spontaneous rodent models of type 2 diabetes, female rodents are protected against hyperglycemia unless they are ovariectomized, and E2 perfusion reverses diabetes in male rodents. Finally, the study of transgenic mice and mice with genetic alteration of E2 secretion or E2 action has shed light on the antidiabetic properties of E2 at a tissue-specific level. Thus, E2 secretion and action in rodents seems to be implicated 1) in adipose tissue biology and the prevention of obesity, 2) in the stimulation of liver fatty acid metabolism and suppression of hepatic glucose production, and 3) in the protection of pancreatic β-cell function/survival and insulin secretion in conditions of oxidative stress.

Anti-cyanobacterial fatty acids released from Myriophyllum spicatum

Abstract  This study was carried out to identify unknown allelochemicals released from Myriophyllum spicatum and to investigate their anti-cyanobacterial effects. A series of analyses of culture solutions and methanol extracts of M. spicatum using gas chromatograph equipped with a mass selective detector revealed that M. spicatum released fatty acids, specifically, nonanoic, tetradecanoic, hexadecanoic, octadecanoic, and octadecenoic acids. Nonanoic, cis-6-octadecenoic, and cis-9-octadecenoic acids significantly inhibited growth of Microcystis aeruginosa, whereas tetradecanoic, hexadecanoic, and octadecanoic acids did not show any effect. When the inhibitory effect of nonanoic acid was compared with those of 4 polyphenols and eugeniin, which are anti-cyanobacterial compounds previously reported to be released by M. spicatum, nonanoic acid was found to be the most inhibitory to M. aeruginosa. These results indicate that not only polyphenols and eugeniin but also fatty acids such as nonanoic acid must be studied to reveal how M. spicatum exerts its allelopathic effect on M. aeruginosa.

Antibacterial activity of a bacteriocin-like substance produced by Bacillus sp. P34 that targets the bacterial cell envelope

Abstract  The objective of this study was to investigate the mode of action of BLS P34, a bacteriocin-like substance (BLS) produced by a novel Bacillus sp. strain P34 isolated from the Amazon basin. The effect of the BLS was tested against Listeria monocytogenes, showing a bactericidal effect at 200 AU (activity units) ml−1, while no inhibition of spore outgrowth of Bacillus cereus was observed with a dose of 1,600 AU ml−1. Growth of Escherichia coli and Salmonella Enteritidis was inhibited, but only when the chelating agent EDTA was co-added with the BLS. The effect of BLS P34 on L. monocytogenes was also investigated by Fourier transform infrared spectroscopy. Treated cells showed an important frequency increase in 1,452 and 1,397 cm−1 and decrease in 1,217 and 1,058 cm−1, corresponding assignments of fatty acids and phospholipids. Transmission electron microscopy showed damaged cell envelope and loss of protoplasmic material. BLS P34 was bactericidal to Gram-positive, and also showed inhibitory effect against Gram-negative bacteria. There is evidence that its mode of action corresponds to that of a membrane-active substance. The knowledge about the mode of action of this BLS is essential to determine its effective application as an antimicrobial agent.

Antiatherogenic components of olive oil

Abstract  Olive oil is the principal source of fat in the Mediterranean diet, which has been associated with a lower incidence of coronary heart disease and certain cancers. Olive oil is characterized by a high proportion of monounsaturated oleic acid, but the main peculiarity of extra-virgin oil is the presence of remarkable quantities of phenolic compounds, notably hydroxytyrosol and oleuropein, that provide high stability and strong taste. Recently, several studies have demonstrated that olive oil phenolics are powerful antioxidants, both in vitro and in vivo, and exert additional potent biologic activities that could partially account for the observed cardioprotective effects of the Mediterranean diet.

Antiarrhythmic and electrophysiological effects of long-chain ω-3 polyunsaturated fatty acids

Abstract  Recent studies indicate that a diet enriched in -3 polyunsaturated fatty acids may prevent sudden cardiac death. The goal of the present study was to elucidate how -3 polyunsaturated fatty acids such as docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and -linolenic acid (ALA; 1–20 M) may affect the cardiac activation and repolarization pattern. For this reason, DHA, EPA or ALA was infused in spontaneously beating isolated rabbit heart (Langendorff technique) and subjected to 256 electrodes epicardial mapping. All compounds exhibited a negative inotropic and chronotropic effect. EPA and ALA, but not DHA, prolonged QTc. The dispersion was enhanced at higher concentrations (>5 M) by DHA and less (or not affected) by the others. The total activation time, reflecting ventricular conduction, was prolonged predominantly by DHA and to a lower extent by the other drugs. Atrioventricular conduction time was slowed only by DHA and EPA. To analyze of the pattern of activation, we determined the timepoint of activation as t(dU/dtmin) for all 256 electrodes. The beat-to-beat similarity of these patterns was moderately reduced by all drugs. Regarding antiarrhythmic activity we found that the threshold for elicitation of a ventricular extrasystole was concentration-dependently enhanced by DHA and EPA, but not by ALA. DHA dose-dependently reduced longitudinal propagation velocity VL and to a lower extent transverse velocity VT. Anisotropy was not significantly changed. EPA and ALA did not exhibit a systematic effect on VL or VT. These results clearly demonstrate that DHA, EPA, and ALA exhibit direct electrophysiological effects with different profiles.

Anthropometry, carbohydrate and lipid metabolism in the East Flanders Prospective Twin Survey: heritabilities

Abstract Aims/hypothesis  We determined the genetic contribution of 18 anthropometric and metabolic risk factors of type 2 diabetes using a young healthy twin population.

Anthropometric features and body composition of young athletes practicing karate at a high and medium competitive level

Abstract.   The aim of the study was to examine the anthropometricfeatures and body composition of athletes practising karate at ahigh and medium competitive level. Our study was carried out ona sample of 35 subjects practising karate and aged from 16.0 to32.5 years. This sample was divided into two groups: group 1(n=14 elite athletes) andgroup 2 (n=21 amateurathletes). Various anthropometric measurements were taken(weight, height both standing and sitting, diameters,circumferences and skinfold thickness) from which differentanthropometric indices were calculated (body mass index, Scelicand Grant indices, arm muscle circumference and area), and thesomatotype was then determined. The body composition of eachsubject was assessed using the skinfold technique and theJackson-Pollock (J-P) and Sloan-Weir (S-W) equations. The twogroups of athletes showed very similar measurements regardinganthropometric characteristics. Only the Scelix index presenteda significantly different value in the two groups (49.6±1.3 forgroup 1 vs. 51.1±1.3 for group 2; p<0.01). Group 1 showed amesomorphic-ectomorphic somatotype, while the amateur athletespresented a balanced mesomorphic type. Moreover, a lowerpercentage of fat mass was more frequent in the first group(J-P=8.1±2.4%; S-W=8.9±3.3%) than in the second one(J-P=9.8±1.6%; S-W=11.2±3.7%), although the differences betweenthe two groups were not significant. We conclude that group 1 ischaracterized by a slightly prominent vertical development ofthe skeletal frame. This could be an anthropometriccharacteristic that is best suited to meet the specificfunctional requirements of this sport. Moreover, both groups ofathletes are characterized by a low percentage of fat mass,particularly the elite group.

Antagonizing potencies of saturated and unsaturated long-chain free fatty acids to isoflurane in goldfish

Purpose  We have previously reported that myristate, a saturated free fatty acid (FFA) with 14 carbons (C14), antagonizes volatile anesthetics in goldfish. The hydrophobicity and molecular configuration of FFAs may play an important role in the antagonizing effect. To examine their contribution, we investigated the antagonizing potencies of saturated and unsaturated long-chain FFAs in goldfish.Methods  Saturated and monounsaturated FFAs of C14–18 were tested. We determined the anesthetic concentration producing a 50% effect (EC50) of isoflurane in the absence or presence of FFA by observing the escape reaction of goldfish against an electrical stimulus.

Antagonism of croton oil inflammation by topical emu oil in CD-1 mice

Abstract  Emu oil is derived from the emu (Dromaius novaehollandiae), which originated in Australia, and has been reported to have anti-inflammatory properties. Inflammation was induced in anesthetized CD-1 mice by applying 50 μL of 2% croton oil to the inner surface of the left ear. After 2 h, the area was treated with 5μL of emu, fish, flaxseed, olive, or liquified chicken fat, or left untreated. Animals were euthanized at 6 h postapplication of different oils, and earplugs (FP) and plasma samples were collected. Inflammation was evaluated by change in earlobe thickness, increase in weight of EP tissue (compared to the untreated ear), and induction in cytokines interleukin (IL)-1α and tumor necrosis factor-α (TNF-α) in EP homogenates. Al-though reductions relative to control (croton oil) were noted for all treatments, auricular thickness and EP weights were, significantly reduced (−72 and −71%, respectively) only in the emu oil-treated group. IL-1α levels in homogenates of auricular tissue were significantly reduced in the fish oil (−57%) and emu oil (−70%) groups relative to the control group. The cytokine TNF-α from auricular homogenates was significantly reduced in the olive oil (−52%) and emu oil (−60%) treatment groups relative to the control group. Plasma cytokine levels were not changed by croton oil treatment. Although auricular thickness and weight were significantly correlated with each other (r=0.750, P<0.003), auricular thickness but not weight was significantly correlated with cytokine IL-1α (r=0.750, P<0.006) and TNF-α (r=0.690, P<0.02). These studies indicate that topical emu oil has anti-inflammatory properties in the CD-1 mouse that are associated with decreased auricular thickness and weight, and with the cytokines IL-1α and TNF-α.

Ant benefits in a seed dispersal mutualism

Abstract  Myrmecochorous plant seeds have nutrient rich appendages, elaiosomes, which induce some ant species to carry the seeds back to their nest where the elaiosome is consumed and the seed is discarded unharmed. The benefits to plants of dispersal of their seeds in this way have been well documented, but the benefits to the ants from consuming the elaiosomes have rarely been measured and are less clear. Ant benefits from myrmecochory were investigated in a laboratory experiment using the ant Myrmica ruginodis and seeds of Ulex species. To separate the effects of elaiosome consumption on the development of newly produced larvae versus existing larvae, ten ‘Queenright’ colonies containing a queen were compared to ten ‘Queenless’ colonies. Six measures of colony fitness over a complete annual cycle were taken: sexual production, larval weight and number, pupal weight and number, and worker survival. Queenless colonies fed with elaiosomes produced 100.0±29.3 (mean ± SE) of larvae compared to non-elaiosome fed colonies which produced 49.6±19.0; an increase of 102%. Larval weight increased in both Queenright and Queenless colonies. In colonies fed with elaiosomes, larvae weighed 1.02±0.1 mg, but in non-elaiosome fed colonies larvae weighed 0.69±0.1 mg; an increase of 48%. The food supplement provided by Ulex elaiosomes was trivial in energetic terms, under the conditions of an ample diet, suggesting that these effects might be due to the presence of essential nutrients. Chemical analysis of Ulex elaiosomes showed the presence of four essential fatty acids and four essential sterols for ants.

Anoxybacillus rupiensis sp. Nov., a novel thermophilic bacterium isolated from Rupi basin (Bulgaria)

Abstract  Three strains of a novel thermophilic, strictly aerobic, Gram-positive, spore-forming hemo-organotrophic bacterium were isolated from three hot springs in the region of Rupi basin, Bulgaria as producers of amylolytic enzymes. Their 16S rRNA gene sequences (first 500 nucleotides) were very similar (99.8%). Strains were able to ferment a wide spectrum of carbohydrates such as sugars, polyols, and polysaccharides like xylan, glycogen and starch. Optimal growth was observed at 55–58°C, and pH at 6.0–6.5. Phylogenetic analysis of the whole 16S rRNA gene sequence clustered the strain R270T with the representatives of the genus Anoxybacillus and with Geobacillus tepidamans. The G + C content of the genomic DNA was 41.7%. DNA–DNA hybridization analysis revealed low homology with the closest relatives (32.0 mol% homology to Geobacillus tepidamans). Fatty acid profile (major fatty acids iso-C15:0 and iso-C17:0) confirmed the affiliation of the strain to the genus Anoxybacillus. On the basis of the data presented here, we propose that strain R270T, represents a new species of the genus Anoxybacillus for which, we recommend the name Anoxybacillus rupiensis sp. nov. (=DSM 17127T = NBIMCC 8387T). The 16S rRNA gene sequence data of a strain R270T have been deposited in the EMBL databases under the accession number AJ879076.

Anomalous high iodine value of squalene and the impact on iodine values of shark liver oils

Abstract  Squalene has six ethylenic bonds, but the experimental iodine values in two different solvent systems—chloroform and cyclohexane/acetic acid—were 25% higher than the theoretical values. We propose that this results from an additional halogen adding at each of the two terminal ethylenic bonds carrying two methyl groups. In the solvent system of cyclohexane alone, the excess is only 3–4% greater than the theoretical. Mixtures of squalene in seal oil confirmed the additivity of the experimental squalene high iodine value and the seal oil fatty acid iodine value with reasonable accuracy but depended on the skill of the operator in obtaining the titration end point for cyclohexane/acetic acid. This observation has particular relevance for shark liver oils and olive oils.

Anomalous high iodine value of squalene and the impact on iodine values of shark liver oils

Abstract  Squalene has six ethylenic bonds, but the experimental iodine values in two different solvent systems—chloroform and cyclohexane/acetic acid—were 25% higher than the theoretical values. We propose that this results from an additional halogen adding at each of the two terminal ethylenic bonds carrying two methyl groups. In the solvent system of cyclohexane alone, the excess is only 3–4% greater than the theoretical. Mixtures of squalene in seal oil confirmed the additivity of the experimental squalene high iodine value and the seal oil fatty acid iodine value with reasonable accuracy but depended on the skill of the operator in obtaining the titration end point for cyclohexane/acetic acid. This observation has particular relevance for shark liver oils and olive oils.

Ankylosing spondylitis and bone mineral density—what is the ideal tool for measurement?

Abstract  Ankylosing spondylitis (AS) is characterised by chronic inflammation and partial ossification, yet vertebral fractures due to osteoporosis, although common, are frequently unrecognised. The aim of this study was to (1) show the frequency of changes in the progress of osteopenia/osteoporosis in AS depending on duration and stage of the disease and (2) assess the ranking of two different methods of bone density measurement in this clinical pattern. We measured bone density in 84 male and female patients with both dual X-ray absorptiometry (DXA) and single energy quantitative computed tomography (SE-QCT). In the initial and advanced stages of the disease, a high decrease in axial bone density could be verified (DXA: osteopenia in 5% and osteoporosis in 9.2%; SE-QCT: osteopenia in 11.8% and osteoporosis in 30.3%). Peripheral bone density decrease as in osteopenia could be proven in 17.6% by DXA measurement. With SE-QCT, a decrease in vertebral trabecular bone density could already be observed in the initial stage and continued steadily during the course of the disease; cortical bone displayed the same trend up to stages of ankylosis. With DXA, valid conclusions are more likely to be expected in less marked ankylosing stages of AS. In stages of advanced ankyloses in the vertebral region (substantial syndesmophytes), priority should be given to SE-QCT, due to the selective measurement of trabecular and cortical bone. The DXA method often yields values that are too high, and the replacement of vertebral trabecular bone by fatty bone marrow is not usually recorded as standard. There may already be an increased risk of bone fracture in AS in osteopenia on DXA along with an osteoporosis already established on SE-QCT.

Animal models of spontaneous plaque rupture: The holy grail of experimental atherosclerosis research

Abstract  Throughout the history of atherosclerosis research we have sought animal models of the disease process that exhibit high frequencies of the features that make human plaque a clinical risk: plaque rupture, mural thrombosis, and intra-plaque hemorrhage. This type of model is needed to determine the mechanisms by which plaques rupture and to design and test therapeutic interventions for stabilizing plaques. Studies of domestic and exotic animals have shown that most species will spontaneously develop fatty streaks and in some cases atheromatous lesions with sufficient time, but that rupture and thrombosis is exceedingly rare. Even with addition of fat and cholesterol to the diet, lesion development is accelerated but does not increase the frequency with which plaques rupture in most animal models. However, recently we have observed high frequencies of intra-plaque hemorrhage in the innominate/brachiocephalic arteries of older, chow-fed, hyperlipidemic, apolipoprotein E-deficient mice, and high frequencies of plaque rupture with mural thrombus in younger apolipoprotein E-deficient mice fed a high-fat diet. This suggests that plaque rupture and secondary thrombosis are frequent and reproducible occurrences at specific sites in apolipoprotein E-deficient mice, and that the timing and pathobiology of the ruptures are influenced by lipid status in this murine model.

Animal glues in mixtures of natural binding media used in artistic and historic objects: identification by capillary zone electrophoresis

Abstract  Animal glues were often used in historic and artistic objects, e.g. as paint ground, as binders for pigments, or as adhesives. The sources were egg, casein, or different collagens. For restoration and conservation purposes it is important to know which kind of animal glue a museum object contains. Capillary electrophoresis can deliver such information, because it enables differentiation among the three proteinaceous glue classes according to their different amino acid patterns after hydrolysis. This work deals with the most relevant problem in practice, whether this identification is obstructed by the presence of other binders, with which they are mixed in many real samples; in particular, interference from plant gums and drying oils was investigated. Capillary electrophoresis of the hydrolysates (after reaction with 6 mol L–1 HCl) was performed with an acidic background electrolyte consisting of chloroacetic acid (51.9 mmol L–1) adjusted with LiOH to pH 2.26. The underivatised analytes were detected with a contactless conductivity detector. It was found that the constituents of the plant gums (monosaccharides) or drying oils (long-chain fatty acids and short-chain dicarboxylic acids) never interfered with identification of the animal glues, as shown for artificial mixtures of the different binders even at tenfold excess over the animal glue, and for egg tempera samples. The method was used to identify the filling material from a statue from the eighteenth century.

Angiotensin-receptor blocking agents and the peroxisome proliferator-activated receptor-γ system

Abstract  The metabolic syndrome leads to cardiovascular disease and type 2 diabetes mellitus, through multiple risks, such as insulin resistance, dyslipidemia, hyperinsulinemia, and hypertension. It also represents a disorder of partial genetic background as mutations of the peroxisome proliferator-activated receptor-gamma (PPAR-γ). Thiazolidinedione agonists for the PPAR-γ system are effective in control of insulin resistance and diabetes. Telmisartan has a molecular structure that imparts partial agonist properties with the PPAR-γ molecule, which results in reductions in glucose and lipid metabolism. Administration of telmisartan to rats on a high-fat, high-carbohydrate diet leads to reductions in glucose, insulin, and triglyceride levels. The results imply that the ARB agent, telmisartan, could treat both the hemodynamic and metabolic aberrations seen in subjects with the metabolic syndrome, such as insulin resistance, glucose intolerance, and hypertension.

Angiotensin II receptor antagonist reduces urinary liver-type fatty acid-binding protein levels in patients with diabetic nephropathy and chronic renal failure

Angiomyofibroblastoma-like tumor (cellular angiofibroma) in the male inguinal region

Abstract  Angiomyofibroblastoma-like tumor is a rare mesenchymal tumor involving the male genital tract. We report a case of an angiomyofibroblastoma-like tumor that arose in the subcutaneous tissue of the left inguinal region in a 50-year-old man. Ultrasonography of the region demonstrated a well-circumscribed subcutaneous mass. Intralesional fat was revealed on magnetic resonance images. Although these imaging features are nonspecific, radiological findings enable considering the diagnosis of angiomyofibroblastoma-like tumor.

Angiographic follow-up after surgical treatment of intracranial aneurysms

Summary  This study was undertaken to evaluate the long-term angiographic outcome of surgically treated aneurysms. In addition, the incidence of recurrent aneurysms, the fate of residual aneurysms and the de novo formation of aneurysms were evaluated. Moreover, morphological conditions such as dilatation, stenosis or irregularity in temporarily clipped vascular segments were analyzed.166 aneurysms were operated on in 136 patients and these 166 aneurysms subsequently underwent late follow-up angiography. Late angiographic follow-up review was obtained at a mean of 46.6±11.5 months postsurgery (range 36–85 months). Out of the 7 aneurysms with known residua, 5 residual aneurysms were determined as unchanged, 1 residual aneurysm as spontaneous thrombosis and 1 residual aneurysm as enlarged. No recurrent aneurysm was found, however two de novo aneurysms were found. During the surgery of 85 anuerysms, 137 vascular segments clipped temporarily were evaluated in terms of morphological changes in postoperative and long-term angiograms. No morphological changes were determined in any vascular segments.

Angiogenesis in rat aortic rings stimulated by very low concentrations of serum and plasma

Abstract  Arterial ring cultures are evaluated for assay of angiogenic activity in physiological fluids. A simplified image analysis algorithm involving public domain software is used to measure microvessels sprouting from rat aortic rings embedded in fibrin. Low concentrations (>1% v/v) of serum and plasma stimulate angiogenesis primarily from the adventitia in a dose-dependent manner. Half-percent serum is more potent than optimal concentrations of some of the important known growth factors, including platelet derived growth factor, vascular endothelial growth factor and basic fibroblast growth factor. Sera and plasmas obtained from rodent, bovine, porcine or human sources have comparable angiogenic activities. The angiogenic activity is found to be heat stable, non-dialyzable and on gel chromatography is co-eluted with albumin. However, purified albumin and the essential fatty acids linoleic and linolenic acids have no effect on angiogenesis. It is concluded that rat aortic rings sprout microvessels, primarily from the adventitia, when cultured with serum or plasma in concentrations below those that support cell culture. Accordingly, the aortic ring assay is a promising system to support isolation of angiogenic factors from blood and other physiological fluids.

Angiogenesis and marrow stromal cell fates: roles in bone strength

Abstract  In vivo osteogenic responses to anabolic stimuli—whether pharmacological or mechanical—are invariably accompanied by neovascularization. Microvascular endothelial-mesenchymal interactions have emerged that provide mechanistic insight into the roles of angiogenesis in the osteoanabolic response; these interactions resemble the epithelial-mesenchymal signaling that controls tissue morphogenesis during prenatal development. Microvascular smooth muscle cells called pericytes function as multipotent mesenchymal progenitors that contribute to bone, fat, cartilage and smooth muscle formation throughout life. This abbreviated overview recounts progress made in the past decade that highlights the physiological contributions of angiogenesis to bone formation and bone strength. It highlights the need to support research that details the mechanisms whereby angiogenesis, metabolic milieu and mechanical stimuli interact to control marrow stromal cell fate during the postnatal developmental process of aging and the disease processes of musculoskeletal frailty.

Angeborene Stoffwechselerkrankungen

Zusammenfassung  In Österreich und Deutschland stammt ein hoher Anteil (20–50%) der Patienten mit Stoffwechselerkrankungen aus türkischen Migrantenfamilien. Das Risiko, an einer im erweiterten Neugeborenenscreening erfassten Stoffwechselerkrankung zu leiden, ist bei Kindern aus türkischen im Vergleich zu inländischen Familien signifikant höher. Eine Ursache dafür ist u. a. die Tradition der konsanguinen Eheschließung (Kusinenehe) in dieser Bevölkerungsgruppe. Verlaufsdaten von Kindern mit Phenylketonurie weisen darauf hin, dass die Diätcompliance bei türkischen im Vergleich zu österreichischen Kindern schlechter ist. Zwischen türkischen und österreichischen Familien zeigten sich Unterschiede in krankheitsrelevantem Wissen, Bewältigungsstrategien, Glaube an Schicksalhaftigkeit und empfundener Abhängigkeit vom medizinischen Personal. Das Wissen um die Besonderheiten in der Führung von Patienten aus fremden Kulturkreisen, die Einbeziehung von professionellen Dolmetschern in Patientengespräche sowie die aktive Einbindung von Migranten in Selbsthilfegruppen sind zur Verbesserung der medizinischen Versorgung dieser Bevölkerungsgruppe erforderlich.

Angeborene Herzfehlbildung als Todesursache bei einem 7-j?hrigen M?dchen

Zusammenfassung  Die Autoren beschreiben und diskutieren den Fall eines 7-jährigen Mädchens, das wegen einer Tonsillitis zunächst ambulant antibiotisch behandelt wird. Nach einigen Tagen entwickeln sich unter der Therapie Erbrechen und Diarrhöen. Nach der Einweisung in ein Kinderkrankenhaus kommt es kurz nach der Aufnahme zu einem Herz-Kreislauf-Stillstand; die Reanimation bleibt erfolglos. Die Sektion zeigt eine angeborene Herzfehlbildung vor allem der rechten Herzkammer mit basaler trabekulärer Hypertrophie und rechtsseitiger Ausflussbehinderung im Sinne einer subvalvulären Pulmonalstenose. Histologisch imponieren in den inneren Myokardschichten neben einem multifokal irregulären Verlauf hypertrophierter Muskelzellen multiple herdförmige Myokardzellnekrosen in verschiedenen Stadien der Organisation sowie Fetteinschlüsse. Ferner findet sich im subendokardialen Myokard eine Rarefizierung intramuraler Gefäße bei sinusartiger Verlängerung der intertrabekulären Ventrikellichtungen. Die beschriebene Fehlbildung lässt sich am ehesten als primäre Kardiomyopathie mit konsekutiver ventrikulärer non-compaction (VNC) klassifizieren. Es muss von einem schicksalhaften Verlauf einer nicht diagnostizierten Herzfehlbildung ausgegangen werden, die möglicherweise im Rahmen eines Infektes dekompensierte. Unklar bleibt, wie das Kind fast 8 Jahre alt werden konnte, ohne klinisch diagnostiziert zu werden.

Anesthetic management of an extremely obese patient

Abstract  We present the case of a morbidly obese woman, with a body mass index (BMI) of 73.7 kg·m−2, who had a gynecological operation under combined general and epidural anesthesia. The patient's trachea was intubated, using a fiberscope, while she was breathing spontaneously after the intravenous injection of fentanyl and propofol as sedatives. Anesthesia was maintained with intravenous propofol and epidural mepivacaine. When the gynecologist placed a sponge in the abdominal cavity to retract the bowel, the patient experienced severe arterial deoxygenation and mild hypotension, due to massive atelectasis of the left lung. Both oxygenation and perfusion were corrected by the removal of the sponge and with the placement of a pillow under the patient's left shoulder. The atelectasis resulted from compression of the left lung by the fatty mediastinum and by the diaphragm being pushed up by the sponge. The hypotension resulted from impaired venous return and hypoxia. The patient suffered no perioperative complications other than atelectasis and a surgical-site infection. Key factors that contributed to the favorable outcome of this patient included a careful tracheal intubation technique, the choice and dose of anesthetic agents, immediate correction of the factors leading to atelectasis, early ambulation, and prophylaxis for deep vein thrombosis.

Anesthetic management of a pediatric patient on a ketogenic diet

Abstract  There are several specific considerations regarding seizure control during the perioperative period in patients who have been placed on a ketogenic diet (KD). A KD is high in fat and low in protein and carbohydrates and has a long history of use for the treatment of intractable seizures in children. Maintaining therapeutic ketosis and modifying the acid–base balance are particularly important for preventing seizures in patients on a KD. We report changes in the biochemical parameters of a patient with double cortex syndrome who was on a KD and who had been scheduled for the treatment of dental caries under sevoflurane anesthesia and acetate Ringer administration. Inhalation induction with a high concentration of sevoflurane should be reconsidered in view of recent reports describing the epileptogenic potential of sevoflurane.

Ancestral proportions and their association with skin pigmentation and bone mineral density in Puerto Rican women from New York city

Abstract  Hispanic and African American populations exhibit an increased risk of obesity compared with populations of European origin, a feature that may be related to inherited risk alleles from Native American and West African parental populations. However, a relationship between West African ancestry and obesity-related traits, such as body mass index (BMI), fat mass (FM), and fat-free mass (FFM), and with bone mineral density (BMD) in African American women has only recently been reported. In order to evaluate further the influence of ancestry on body composition phenotypes, we studied a Hispanic population with substantial European, West African, and Native American admixture. We ascertained a sample of Puerto Rican women living in New York (n=64), for whom we measured BMI and body composition variables, such as FM, FFM, percent body fat, and BMD. Additionally, skin pigmentation was measured as the melanin index by reflectance spectroscopy. We genotyped 35 autosomal ancestry informative markers and estimated population and individual ancestral proportions in terms of European, West African, and Native American contributions to this population. The ancestry proportions corresponding to the three parental populations are: 53.3±2.8% European, 29.1±2.3% West African, and 17.6±2.4% Native American. We detected significant genetic structure in this population with a number of different tests. A highly significant correlation was found between skin pigmentation and individual ancestry (R2=0.597, P<0.001) that was not attributable to differences in socioeconomic status. A significant association was also found between BMD and European admixture (R2=0.065, P=0.042), but no such correlation was evident with BMI or the remaining body composition measurements. We discuss the implications of our findings for the potential use of this Hispanic population for admixture mapping.

Ancestral Mechanisms in Modern Environments

Abstract  It is commonly assumed that the desire for a thin female physique and its pathological expression in eating disorders result from a social pressure for thinness. However, such widespread behavior may be better understood not merely as the result of arbitrary social pressure, but as an exaggerated expression of behavior that may have once been adaptive. The reproductive suppression hypothesis suggests that natural selection shaped a mechanism for adjusting female reproduction to socioecological conditions by altering the amount of body fat. In modern Western culture, social and ecological cues, which would have signaled the need for temporary postponement of reproduction in ancestral environments, may now be experienced to an unprecedented intensity and duration.

Anatomy of the SMAS Revisited

Anatomy of the smallest coleoptera, featherwing beetles of the tribe nanosellini (Coleoptera, Ptiliidae), and limits of insect miniaturization

Abstract  The internal structure of Nanosella sp., Primorskella sp., and Porophila sp. was described from serial sections and total preparations using light and transmission electron microscopy. The most important structural features related to miniaturization are the absence of midgut muscles, abortion of two Malpighian tubes, a decrease in the number of abdominal spiracles, strong reduction of the tracheal system, the absence of the heart, reduction of the circulatory system and its substitution by the fat body, strong oligomerization and concentration of the nervous system, a decrease in the size and number of neurons, and reduction of the left testis and left ovary. The internal structure of featherwing beetles was analyzed for the first time using 3D computer models, which allowed us to demonstrate changes in the relative volume of organs accompanying the body diminution. The excretory and digestive systems change isometrically, while others change allometrically. The relative volume of the musculature decreases, and that of the nervous and reproductive systems increases. The skeleton mass also increases. The possible factors limiting further diminution in Ptiliidae (egg size, the volume of the reproductive and nervous systems, and the skeleton mass) are discussed.

Anatomophysiologie des algies pudendales

Résumé  Les algies pudendales sont périnéales de type tronculaire et somatique. Elles siègent dans le territoire du nerf atteint et sont positionnelles, ce qui évoque un phénomène de compression lors de la position assise. L’anatomophysiologie de ces douleurs met l’accent sur plusieurs items: a) définition et innervation du périnée; b) étude du parcours des voies de la douleur; étude des structures inhibitrices tout au long de ce trajet. Le périnée: il est embryologiquement situé dans le plan infralévatorien, et contient les organes érecteurs, les sphincters et du tissu graisseux. Il s’étend des organes génitaux à l’anus. Son innervation est mixte: le nerf pudendal issu de S3 représente l’innervation cutanée et musculaire striée. Il contient un important contingent de fibres orthosympathiques qui gagneront la cha?ne latérovertébrale. Le pelvis est supralévatorien, contient les viscères pelviens et n’est innervé que par le système végétatif. Les voies de la douleur sont issues des trois feuillets embryonnaires. Les douleurs extéroceptives (cutanées) gagnent l’apex de la corne grise dorsale; les fibres proprioceptives (muscles, fascias, tendons) gagnent l’isthme; les fibres intéroceptives (viscères) gagnent la base. Des neurones convergents de la lame V de Rexed vont rassembler les informations douloureuses dans le tractus spinothalamique ventral (STV) et dorsal (STD). Dans le tronc cérébral, le STD monte à petite vitesse et va ensuite gagner le thalamus. De nombreuses fibres vont être stoppées dans la formation réticulée. Le STV devient satellite de la voie lemniscale et gagne le thalamus à grande vitesse avec elle. Dans le noyau VPL du thalamus, les fibres du STD vont projeter dans le cortex préfrontal et donner à la douleur sa dimension qualitative. Le STV arrivé le premier projette immédiatement dans le gyrus postcentral pour localiser la douleur. Parallèlement, l’hypothalamus, informé par la réticulée, accompagne le sentiment douloureux d’un cortège hormonal réactionnel. Le cortex limbique puisant ses informations du néocortex stockera ou non la douleur en mémoire, en l’accompagnant d’une composante émotionnelle. Les voies de la douleur viscérale sont semblables à celles des douleurs somatiques dans le système nerveux central. La connaissance de leur composante extra-axiale permet d’orienter les infiltrations (rameaux communicants). Les structures inhibitrices sont à tous les niveaux: gate control dans la moelle spinale, filtre réticulaire et thalamique, systèmes inhibiteurs diffus et néocortex moteur. Leur connaissance oriente vers les techniques de neurostimulation quand le traitement étiopathologique échoue. Au total, devant une douleur périnéale, la connaissance anatomique doit faire évoquer parfois une pathologie tronculaire somatique et abandonner le réflexe de rendre responsables des structures viscérales n’appartenant pas au périnée, et devenant victimes d’un comportement thérapeutique inadapté.

Anatomie und Einteilung der Rotatorenmanschettendefekte

Zusammenfassung  Defekte an der Rotatorenmanschette müssen anatomisch genau beschrieben und die zugrunde liegende Ursache muss erkannt werden, um eine sinnvolle und einheitliche Therapie zu ermöglichen. Die bildgebenden Verfahren und besonders die Magnetresonanztomographie erlauben neben einer ersten Abschätzung des Sehnenschadens v. a. die Beurteilung einer möglichen Atrophie und Verfettung der zugehörigen Muskulatur. Eine exakte Klassifikation v. a. kleinerer oder teilweiser Rotatorenmanschettenläsionen gelingt aber nur mit der arthroskopischen Beurteilung des artikularseitigen wie bursaseitigen Sehnenansatzes. Solch kleinere Schäden der Rotatorenmanschette können heute in arthroskopischer Technik auch behandelt werden. Sofern ein größerer Defekt arthroskopisch nicht zu verschließen ist, kann zumindest seine Rekonstruierbarkeit beurteilt werden, sodass sich vergebliche offene Eingriffe vermeiden lassen.

Anatomical pitfalls during encircling cryoablation of the left atrium for atrial fibrillation therapy in the pig

Abstract  We previously published encircling endocardial cryo-isolation of the pulmonary vein (PV) region. This study documented mechanisms of isolation failure using CARTO? mapping. Cryo-isolation used a modified Surgifrost? introduced via a Universal Cardiac Introducer? on the left atrial appendage. Of five pigs, two had incomplete isolation and repeat mapping: Activation was over Bachmann’s bundle (BB) in one and the coronary sinus (CS) in the other. Repeat cryoablation failed to eliminate gaps. Histologically, the BB gap had nonlesioned sub-epicardial fibres and thick fat covering the cryolesioned BB: fat protecting the epicardium from cryoablation. The inferior gap had a large CS, and a thick myocardium bridging the isthmus: myocardial thickness and CS thermal sink preventing transmural cryolesions. CARTO? mapping localized gaps. Although the CS is known to cause failure, its protective mechanism is not well documented. The BB gap is novel. These findings have important clinical implications for isolation of the PV region.

Anatomical bases of the surgical dissection of the interatrial septum: a morphological and histological study

Abstract Background  The interatrial septum (IAS) can be dissected to resect pulmonary tumors invading the left atrium. The aim of this study was to describe the dissected structures, and to expose the benefits, the limits, and the embryologic reasons of such dissection.

Anatomical bases of prolonged ilio-inguinal-hypogastric regional anesthesia

Abstract  Although anesthesia and post-operative analgesia are associated with specific morbidity, regional anesthesia is not systematically given during groin hernia surgery. The goals of this work were to determine the anatomical bases of safe ilio-inguinal (II)-hypogastric anesthesia that can be prolonged into the post-operative period and to validate this technique on anatomical preparations and in clinical situations. We studied the courses of the ilio-hypogastric (IH) and II nerves in 33 halves of 20 embalmed adult cadavers. The intermediate portion of the IH and II nerves, located between the transverse and the internal oblique muscles, were found to be suitable for a simultaneous block with a single injection. We assessed the feasibility of injecting a percutaneous infiltration into this space by injecting a dye before dissection. In 75% of cases, we observed percutaneous coloring of the nerves, confirming that this site was suitable. To guide the infiltration, the points where the nerves passed through the transverse and the internal oblique muscles were located from the iliac crest and anterior and superior iliac spine, respectively. The nerve trunks were grouped for over 5 cm in a cell-fat layer running between these two deep muscles. It was possible to position a micro-catheter in this anatomical space to allow repeated injections. The results of this anatomical study were used to modify the technique so that it could be used to provide regional anesthesia in five patients operated on for hernia. Post-operative pain was very effectively controlled in four cases with no complications.

Anatomical and molecular reinvestigation of lamprey endostyle development provides new insight into thyroid gland evolution

Abstract  The thyroid gland of vertebrates is considered to be homologous to the endostyle of non-vertebrate chordates (cephalochordates, urochordates), a key character for understanding the origin and evolution of the chordate body plan. In lampreys, the larval endostyle transforms into an adult thyroid gland during metamorphosis, reflecting evolutionary changes that occurred in the vertebrate lineage. Focussing on thyroid-like cells in the endostyle, we here relate morphologically visible steps of lamprey (Lampetra fluviatilis) endostyle differentiation to embryonic stages and determine the onset of thyroid-like function. Analysing lamprey endostyle development using semi-thin histological sections, immunohistochemical detection of thyroid hormone, and the molecular marker thyroid transcription factor1 (Ttf1) refines our current view of the homology between endostyle and thyroid gland. In contrast to earlier literature, we find that a duct always persists to connect the endostyle lumen to the pharynx, a structure that resembles the thyroglossal duct in thyroid development and could further support the homology between endostyle and thyroid. Before the onset of thyroid-like function, Ttf1 expression becomes restricted to the ventral part of the endostyle, on the one hand showing that dorsal thyroid-like cells produce thyroid hormone in the absence of Ttf1, and on the other suggesting that Ttf1 was initially involved in specifying ventral fates in the endostyle.

Anatomic and Physiologic Advantages of Totally Subfacial Breast Implants

Abstract  For the past 4 years, whenever it was possible to choose between the subglandular or the submuscular location for the implant pocket (in cases with no precise indication for the submuscular location), the authors have opted for the totally subfascial plane (subaponeurotic) to avoid the disadvantages of the other locations and to obtain additional benefits. The subfascial plane lies below the deep thoracic fascia, or deep aponeurosis, which is not only that of the pectoralis (the muscle connecting with the breast in its upper two-thirds only). The implant is placed completely beneath this fascia, which covers, in addition to the pectoralis major muscle, the serratus, the lateral oblique, and the rectus anterior muscles. This study included 100 women who had breast implanted in the subfascial (subaponeurotic) plane. Of these women, 63 were thin patients with little fatty tissue. Textured surface implants were used McGhan style 120 and 110. All procedures were approached through an inferior periareolar incision. Excellent coverage of the implant as well as natural shape and mobility was achieved for all patients. No rippling in the upper half was observed. Two patients had Baker grade 2 capsular contracture (2%). No seromas or infections were seen, and only once was a surgical exploration necessary for excess drainage volume (1%). In the immediate postoperative period, less edema was recorded and recovery was faster than with the other two procedures: submuscular and subglandular. When circumstances indicate a subglandular pocket, the subfascial plane would seem to be the logical place for mammary implants.

Anaplerotic roles of pyruvate carboxylase in mammalian tissues

Abstract.  Pyruvate carboxylase (PC) catalyzes the ATP-dependent carboxylation of pyruvate to oxaloacetate. PC serves an anaplerotic role for the tricarboxylic acid cycle, when intermediates are removed for different biosynthetic purposes. In liver and kidney, PC provides oxaloacetate for gluconeogenesis. In adipocytes PC is involved in de novo fatty acid synthesis and glyceroneogenesis, and is regulated by the peroxisome proliferator-activated receptor-γ, suggesting that PC is involved in the metabolic switch controlling fuel partitioning toward lipogenesis. In islets, PC is necessary for glucose-induced insulin secretion by providing oxaloacetate to form malate that participates in the ‘pyruvate/malate cycle’ to shuttle 3C or 4C between mitochondria and cytoplasm. Hyperglycemia and hyperlipidemia impair this cycle and affect glucose-stimulated insulin release. In astrocytes, PC is important for de novo synthesis of glutamate, an important excitatory neurotransmitter supplied to neurons. Transcriptional studies of the PC gene pinpoint some transcription factors that determine tissue-specific expression.

Anaplerotic molecules: Current and future

Summary  This review presents the concepts of anaplerosis and cataplerosis in relation to the regulation of citric acid cycle operation. Anaplerosis is the re-filling of the catalytic intermediates of the cycle that carry acetyl-CoA as it is oxidized. The main anaplerotic substrates are pyruvate, glutamine/glutamate and precursors of propionyl-CoA (odd-chain fatty acids, specific amino acids, C5-ketone bodies). Cataplerosis balances anaplerosis by removing excess intermediates from the citric acid cycle. The properties of the main anaplerotic substrates are reviewed from the point of view of potential clinical applications to the treatment of some inherited and acquired conditions.

Anaplerotic diet therapy in inherited metabolic disease: Therapeutic potential

Summary  Beginning with phenylketonuria, dietary therapy for inborn errors has focused primarily on the restriction of the precursor to an affected catabolic pathway in an attempt to limit the production of potential toxins. Anaplerotic therapy is based on the concept that there may exist an energy deficit in these diseases that might be improved by providing alternative substrate for both the citric acid cycle (CAC) and the electron transport chain for enhanced ATP production. This article focuses on this basic problem, as it may relate to most catabolic disorders, and provides our current experience involving inherited diseases of mitochondrial fat oxidation, glycogen storage, and pyruvate metabolism using the anaplerotic compound triheptanoin. The observations have led to a realization that ‘inter-organ’ signalling and ‘nutrient sensors’ such as adenylate monophosphate mediated-protein kinase (AMPK) and mTOR (mammalian target of rapamycin) appear to play a significant role in the intermediary metabolism of these diseases. Activated AMPK turns on catabolic pathways to augment ATP production while turning off synthetic pathways that consume ATP. Information is provided regarding the inter-organ requirements for more normal metabolic function during crisis and how anaplerotic therapy using triheptanoin, as a direct source of substrate to the CAC for energy production, appears to be a more successful approach to an improved quality of life for these patients.

Analyzing seed weight, fatty acid composition, oil, and protein contents in Vernonia galamensis germplasm by near-infrared reflectance spectroscopy

Abstract   Vernonia galamensis is a potential new industrial oilseed crop from the Asteraceae family. The interest in this species is due to the presence of a high vernolic acid content of its seed oil, which is useful in the oleochemical industry for paints and coatings. The development of a rapid, precise, robust, nondestructive, and economical method to evaluate quality components is of major interest to growers, processors, and breeders. NIR reflectance spectroscopy (NIRS) is routinely used for the prediction of quality traits in many crops. This study was conducted to establish a rapid analytical method for determining the quality of intact seeds of V. galamensis. A total of 114 Vernonia accessions were scanned to determine seed weight, FA composition, oil, and protein contents using NIRS. Conventional chemical analysis for FA composition, total oil, and protein contents were performed by GC, Soxhlet extraction, and the Dumas combustion method, respectively. Calibration equations were developed and tested through cross-validation. The coefficient of determination in cross-validation for FA ranged from 0.47 (linoleic acid) to 0.55 (vernolic acid), and for oil, protein, and seed weight from 0.71 (oil) to 0.86 (seed protein). It was concluded that NIRS calibration equations developed for seed weight and seed quality traits can be satisfactorily used as early screening methods in V. galamensis breeding programs.

Analyzing biodiesel: standards and other methods

Abstract  Biodiesel occupies a prominent position among the alternatives to conventional petrodiesel fuel owing to various technical and economic factors. It is obtained by reacting the parent vegetable oil or fat with an alcohol )transesterification) in the presence of a catalyst to give the corresponding monoalkyl esters, which are defined as biodiesel. Because of the nature of the starting material, the production process, and subsequent handling, various factors can influence biodiesel fuel quality. Fuel quality issues are commonly reflected in the contaminants or other minor components of biodiesel. This work categorizes both the restricted species in biodiesel and the physical properties prescribed by the standards, and details the standard reference methods to determine them as well as other procedures. Other aspects of biodiesel analysis, including production monitoring and assessing biodiesel/petrodiesel blends, are also addressed. The types of analyses include chromatographic, spectroscopic, physical properties-based, and wet chemical methods. The justifications for specifications in standards are also addressed.

Analytical supercritical fluid extraction with lipase catalysis: Conversion of different lipids to methyl esters and effect of moisture

Abstract  The fat content of lipid-containing samples has been determined by extraction of the fat with supercritical carbon dioxide, followed by enzyme-catalyzed methylation of the fat under supercritical conditions, prior to gas chromatography (GC) analysis. This study was initiated to determine the effect of moisture content on the extraction and conversion of lipids in oilseed and meat samples to their fatty acid methyl ester (FAME) derivatives. These samples were freeze-dried or mixed with Hydromatrix and compared with untreated control samples by employing the above-described supercritical fluid extraction-reaction sequence. Particular attention was focused on minor constituents, such as phospholipids and cholesteryl esters, to see if they could be extracted and derivatized by the above technique. Recoveries and reaction conversions of the lipid species were determined with the aid of GC, high-performance liquid chromatography, and supercritical fluid chromatography for analyses of the extracted lipids. Total fat values were higher from the freeze-dried meat and oilseed samples than from samples mixed with Hydromatrix or left untreated. Extraction of cholesteryl esters was better than 90%, and conversion of the cholesteryl esters to FAME was 93% or higher. Extraction of phosphatidic acid was only 88% compared to more than 90% recoveries for the other phospholipid species. FAME conversion was better than 96% for all phospholipid samples in the study.

Analytical methods for the determination of persistent ingredients of personal care products in environmental matrices

Abstract  Concern about the environmental fate and potential effects of synthetic organic chemicals used in soaps, lotions, toothpaste, and other personal care products continues to increase. This review describes procedures used for the analysis of five classes of these compounds–synthetic musk fragrances, antimicrobials, ultraviolet filters, insect repellents, and parabens–in water, sediment, sewage sludge, air, and aquatic biota. The primary focus is on sample extraction and preparation methods for these compounds. Instrumental methods commonly used for these compounds are also discussed.

Analytical characterization of the persistent residues after microbial degradation of mineral oils

Abstract  The residual fractions remaining after microbial degradation of diesel fuel, different deparaffinized raffinates and extracts from long-term contaminated soils were analyzed by liquid chromatography, gas chromatography, infrared spectrometry and mass spectrometry. The quantity of saturated hydrocarbons decreased after the microbial treatment, whereas the portion of polar compounds increased. The total content of aromatics changed only insignificantly. n-Paraffins < C26 were found to be no longer present in mineral oils degraded to exhaustion. Infrared spectrometry revealed oxygen compounds in the residues, mainly ketones, fatty acids and esters. Elementary analysis confirms the presence of nitrogen, oxygen and sulphur compounds in the degraded products. The gas chromatograms of high boiling oils, as well as of residues and extracts, consist mainly of a large base envelope (about 95% of the total area); thus gc/ms coupling reaches the limits of its applicability. However, mass spectrometry with direct inlet gives valuable information regarding hydrocarbon type analysis. The results revealed the preferable degradation of alkanes, 1-ring aliphatics and benzenes and an enrichment of condensed cycloaliphatics and aromatics. The latter compounds are known to be resistant to microbial attack.

Analytical characteristics of virgin olive oil from young trees (arbequina cultivar) growing under linear irrigation strategies

Abstract  The effect of a linear (vs. effective crop coefficient, K c), irrigation strategy applied to young olive trees (Arbequina cv.) on the qualitative and quantitative parameters of virgin olive oil quality was studied. Although linear irrigation strategy did not affect the quality indexes used to classify olive oil by commercial grades, it did influence other important parameters such as total phenol content, bitter index oxidative stability, and the sensorial appraisal. All of these of olive oil qualities were negatively associated with the amount of applied irrigation water. Pigment content of oils determined by carotenoid and chlorophylls also was negatively associated with the amount of water supplied. No consistent relation was found for α-tocopherol and fatty acid content of olive oil in relation to these treatments.

Analysis, fate studies and monitoring of the antifungal agent clotrimazole in the aquatic environment

Abstract  The analysis and presence of clotrimazole, an antifungal agent with logK OW > 4, was thoroughly studied in the aquatic environment. For that reason analytical methods based on gas chromatography–mass spectrometry and liquid chromatography–tandem mass spectrometry were developed and validated to quantify clotrimazole with limits of quantification down to 5 and 1 ng/L, respectively. Both methods were compared in an intercalibration exercise. The complete mass-spectrometric fragmentation pattern could be elucidated with the aid of quadrupole time of flight mass spectrometry. Since clotrimazole tends to adsorb to laboratory glassware, studies on its adsorption behaviour were made to ensure the appropriate handling of water samples, e.g. pH, storage time, pretreatment of sampling vessels or material of the vials used for final extracts. The phenomena of adsorption to suspended matter were investigated while analysing different waste-water samples. Application of the methods in various investigated wastewater and surface water samples demonstrated that clotrimazole could only be detected in the low nanogram per litre range of anthropogenic influenced unfiltered water samples after acidification to pH 2.

Analysis of whole cellular fatty acids and anastomosis relationships of binucleate Rhizoctonia spp. associated with Ceratobasidium cornigerum

Abstract  Previous research has demonstrated that whole cellular fatty acids analysis is a useful tool for identifying and establishing taxonomic relationships between anastomosis groups (AGs) and related Rhizoctonia isolates. In this experiment, the composition of fatty acid of 28 isolates of teleomorph genus Ceratobasidium cornigerum, consisting of binucleate Rhizoctonia, AG-A, AG-B(o), AG-C, AG-P, and AG-Q, was evaluated using gas chromatography. Eleven fatty acids identified, i.e., myristic, pentadecanoic, palmitic, 2-hydroxypalmitic, palmitoleic, heptadecanoic, 9-heptadecenoic, stearic, oleic, linoleic, and linolenic acids, were present in isolates of AG-A, AG-B(o), AG-C, AG-P, and AG-Q. The major fatty acids, palmitic, oleic, and linoleic acids, were common in all isolates, constituting 87.1% to 94.7% of the whole cellular fatty acids identified. Isolates within the same AG were closely clustered, whereas isolates from different AGs were clearly and distinctly clustered based on average linkage cluster analysis of whole cellular fatty acids. Principal-component analysis generated from all fatty acids also confirmed the divergent separation of the 5 AGs of binucleate Rhizoctonia.

Analysis of transcripts in methyl jasmonate-treated ginseng hairy roots to identify genes involved in the biosynthesis of ginsenosides and other secondary metabolites

Abstract  Methyl jasmonate (MeJA) treatment increases the levels of plant secondary metabolites, including ginsenosides, which are considered to be the main active compounds in ginseng (Panax ginseng C.A. Meyer). To create a ginseng gene resource that contains the genes involved in the biosynthesis of secondary metabolites, including ginsenosides, we generated 3,134 expression sequence tags (ESTs) from MeJA-treated ginseng hairy roots. These ESTs assembled into 370 clusters and 1,680 singletons. Genes yielding highly abundant transcripts were those encoding proteins involved in fatty acid desaturation, the defense response, and the biosynthesis of secondary metabolites. Analysis of the latter group revealed a number of genes that may be involved in the biosynthesis of ginsenosides, namely, oxidosqualene cyclase (OSC), cytochrome P450, and glycosyltransferase. A novel OSC gene was also identified by this analysis. RNA gel blot analysis confirmed that transcription of this OSC gene, along with squalene synthase (SS) and squalene epoxidase (SE) gene transcription, is increased by MeJA treatment. This ginseng EST data set will also provide important information on the genes that are involved in the biosynthesis of other secondary metabolites and the genes that are responsive to MeJA treatment.

Analysis of TNT (2,4,6-Trinitrotoluene)-Inducible Cellular Responses and Stress Shock Proteome in Stenotrophomonas sp. OK-5

Abstract  In this study, the cellular responses of Stenotrophomonas sp. OK-5 to explosive 2,4,6-trinitrotoluene (TNT) have been extensively analyzed. The stress shock proteins, which might contribute to enhancing cellular resistance to TNT-mediated toxicity, were induced at different concentrations of TNT used as a substrate for cell culture of Stenotrophomonas sp. OK-5 capable of utilizing TNT. Proteomic analysis for 2-DE of soluble protein fractions from the culture of OK-5 exposed to TNT demonstrated approximately 300 spots on the silver-stained gel ranging from pH 3 to pH 10. Among them, 10 spots significantly induced and expressed in response to TNT were selected and analyzed. As the result of internal amino acid sequencing with ESI-Q TOF mass spectrometry, TNT-mediated stress shock proteins such as DnaK, OmpW, and OsmC were identified and characterized. Survival of strain OK-5 was periodically monitored in the presence of different concentrations of TNT along with the production of the stress shock proteins. Cells of strain OK-5 pre-exposed to TNT had in improved survival tolerance. Analysis of total cellular fatty acids in strain OK-5 suggested that several saturated or unsaturated fatty acids might increase or decrease under TNT-mediated stress condition. Scanning electron microscopy of cells treated with 0.8 mM TNT for 12 h revealed irregular rod shapes with wrinkled surfaces.

Analysis of the seed oil of Heisteria silvanii (Olacaceae)—A rich source of a novel C18 acetylenic fatty acid

Abstract  Besides some usual fatty acids (FA), two conjugated ene-yne acetylenic FA [trans-10-heptadecen-8-ynoic acid (pyrulic acid) (7.4%), and trans-11-octadecen-9-ynoic acid (ximenynic acid) (3.5%)], a novel ene-yne-ene acetylenic FA [cis-7, trans-11-octadecadiene-9-ynoic acid (heisteric acid) (22.6%)], and 9,10-epoxystearic acid (0.6%) could be identified in the seed oil of Heisteria silvanii (Olacaceae). Two further conjugated acetylenic FA [9,11-octadecadiynoic acid (0.1%) and 13-octadecene-9,11-diynoic acid (0.4%)] were identified tentatively by their mass spectra. The FA mixture has been analyzed by gas chromatography/mass spectrometry (GC/MS) of their methyl ester and 4,4-dimethyloxazoline derivatives. The structure of heisteric acid was elucidated after isolation via preparative silver ion thin-layer chromatography and by various spectroscopic methods [ultraviolet; infrared; 1H, 13C nuclear magnetic resonance (NMR); 1H−1H and 1H−13C correlation spectroscopy]. To determine the position of the conjugated ene-yne-ene system, the NMR spectra were also measured after addition of the lanthanide shift reagent Resolve-Al EuFODTM. Furthermore, the triyglyceride mixture was analyzed by high-temperature GC and high-temperature GC coupled with negative chemical ionization MS. A glass capillary column coated with a methoxy-terminated 50%-diphenyl-50%-dimethylpolysiloxane was used for the separation of the triacylglycerol (TAG) species. No evidence of decomposition of the TAG species containing conjugated ene-yne-ene FA was observed. Twenty-six species of the separated TAG were identified by means of their abundant quasi molecular ion [M−H]− and their corresponding carboxylate anions [RCOO]− of the fatty acids, respectively. The major molecular species of the TAG were found to be 16:0/18:1/18:1, 16:0/18:1/18:3 (heisteric acid), 17:2 (pyrulic acid)/18:1/18:1, 18:1/18:1/18:3 (heisteric acid). The TAG containing acetylenic FA showed an unexpected increase of the retention time in comparison to the TAG containing usual FA, thus making the prediction of the elution order of lipid samples containing acetylenic FA difficult.

Analysis of the fatty oil of Pistacia eurycarpa nuts by gas chromatography/mass spectrometry

Analysis of the Active Compounds in Different Parts of the Schisandra chinensis Plant by Means of Pyrolysis-GC/MS

Summary.  Different parts of the S. chinensis tree (seeds, seed shells, fruits, leaves, and shoots) were characterized by means of analytical pyrolysis – gas chromatography/mass spectrometry. The samples were pyrolyzed at 350°C leading to the evaporation of the thermally stable lignans. Besides the quantification of the lignans deoxyschisandrin, gomisin N, schisandrin, wuweizisu C, gomisin A, and angeloylgomisin H, further information about the composition of the plant parts, such as lignin, terpene, fatty acid, and carbohydrate content, could be obtained. The results were compared to the ones obtained by supercritical fluid extraction with carbon dioxide as well as literature data and were found to match.

Analysis of tensides in complex samples with comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry

Abstract  Multidimensional gas-chromatographical analysis of various tensides of natural or synthetic origin in cosmetic products is demonstrated. Comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry allows the qualitative and quantitative determination of alkyl polyglucosides (AG), fatty alcohol ethoxylates (FAEO), fatty alcohol sulfates (FAS), fatty alcohol ether sulfates (FAES) and cocamidopropyl betaines (CAPB) in shower gel and cleaning agents. The samples were aliquoted in two parts. The first part was silylated, diluted and analysed; then, in order to detect anionic tensides (FAES, FAS) too, the second aliquot was hydrolysed before being silylated for analysis. Because of their amphoteric character, the betaines can only be analysed by gas chromatography after thermal decomposition in the injector, which leads to the corresponding amidoamines among other products.

Analysis of synovial fluid from clinically healthy Iranian fat-tailed sheep

Abstract  In this study synovial fluid from the radiocarpal joints of 100 clinically healthy Iranian sheep (Lori-Bakhtiari) were analyzed. Total nucleated cell count (TNCC) of the synovial fluid was 178.9±75 cells/l (mean±SD). Lymphocytes were the predominant cell type composing 48.34±17.2% of the cells found in the synovial fluid, whereas monocytes, macrophages and neutrophils composed 36.52±3.5%, 12.75±5.9% and 2.28±1.18% of the cells found in the synovial fluid, respectively. The glucose concentration of synovial fluid was 44.9±9 mg/dl. The concentration of total protein, albumin and globulin of the synovial fluid were 2.31±0.55, 1.49±0.38, 0.81±0.28 g/dl, respectively. The albumin to globulin ratio (A/G) was 2.02±0.61. Age and sex had no significant effects on TNCC, percentage of lymphocytes, monocytes, macrophages and neutrophils, concentration of total protein, albumin, globulin, and A/G ratio of fluid from the radiocarpal joint. However, glucose concentration in radiocarpal fluid in sheep less than 1-year-old was significantly (P0.05) higher (48.27±1.4 mg/dl) than 1- to 2-years-old (42.1±1.4 mg/dl) and more than 2-years-old sheep (43.9±1.8 mg/dl). No significant differences were found between right and left limbs for any parameters evaluated in this study.

Analysis of seed oil from ricinus communis and dimorphoteca pluvialis by gas and supercritical fluid chromatography

Abstract  The seed oils from Dimorphoteca pluvialis and Ricinus communis contain hydroxy fatty acids. Dimorphoteca pluvialis contains Δ-9-hydroxy-10t, 12t-octadecadienoic acid (dimorphecolic acid) and R. communis contains Δ-12-hydroxy-9c-octadecenoic acid (ricinoleic acid). The oils were derivatized and analyzed to determine the content of hydroxy fatty acids. The trimethylsilyl fatty acid methyl ester (TMS-FAME) derivatives were analyzed by capillary gas chromatography (GC), and the free fatty acid (FFA) derivatives and the oils were analyzed by capillary supercritical fluid chromatography (SFC). Further, mass spectroscopy of the TMS-FAME derivatives was performed to check the purity of the derivatives. The results from the GC analyses of TMS-FAME corresponded to the results found by SFC analysis of the FFA. The content of ricinoleic acid in the glycerolipids of R. communis was 87.7 wt%, and the content of dimorphecolic acid in D. pluvialis was 54.0 wt%. The methods were evaluated with respect to the cost, ease, and time needed for sample preparation and analysis.

Analysis of nutrient components of food for Asian elephants in the wild and in captivity

Abstract  Thirty-seven wild plants as food for Asian elephants in the field in Simao, Yunnan province, China and five cultivated plants as food for captive elephants in the Beijing Zoo were collected and analyzed for their main nutrient components. Protein, fat, fiber, dry material, ash as well as major microelements: calcium, kalium, zincum, sodium in the food were analyzed by standard methodology. No significant differences were found between the wild plants taken in the field and forage provided in captivity. However, the calcium content in the forage is significantly less than the average of those in the wild plants. It is suggested that the increase in calcium intake may contribute to the relief of low plasma calcium diseases of elephants in captivity.

Analysis of novel hydroperoxides and other metabolites of oleic, linoleic, and linolenic acids by liquid chromatography-mass spectrometry with ion trap MSn

Abstract  Linoleate is oxygenated by manganese-lipoxygenase (Mn-LO) to 11S-hydroperoxylinoleic acid and 13R-hydroperoxyoctadeca-9Z,11E-dienoic acid, whereas linoleate diol synthase (LDS) converts linoleate sequentially to 8R-hydroperoxylinoleate, through an 8-dioxygenase by insertion of molecular oxygen, and to 7S,8S-dihydroxylinoleate, through a hydroperoxide isomerase by intramolecular oxygen transfer. We have used liquid chromatography-mass spectrometry (LC-MS) with an ion trap mass spectrometer to study the MSn mass spectra of the main metabolites of oleic, linoleic, α-linolenic and γ-linolenic acids, which are formed by Mn-LO and by LDS. The enzymes were purified from the culture broth (Mn-LO) and mycelium (LDS) of the fungus Gaeumannomyces graminis. MS3 analysis of hydroperoxides and MS2 analysis of dihydroxy- and monohydroxy metabolites yielded many fragments with information on the position of oxygenated carbons. Mn-LO oxygenated C-11 and C-13 of 18∶2n−6, 18∶3n−3, and 18∶3n−6 in a ratio of ∼1∶1–3 at high substrate concentrations. 8-Hydroxy-9(10)expoxystearate was identified as a novel metabolite of LDS and oleic acid by LC-MS and by gas chromatography-MS. We conclude that LC-MS with MSn is a convenient tool for detection and identification of hydroperoxy fatty acids and other metabolites of these enzymes.

Analysis of nonvolatile lipid oxidation products in vegetable oils by normal-phase high-performance liquid chromatography with mass spectrometric detection

Abstract  Nonvolatile triacylglyceride (TAG) oxidation products play an important role in the oxidative degradation of lipids. They serve as a reservoir of oxygen-containing species and hence can act as off-flavor precursors or as initiators for further oxidation reactions. Possible nonvolatile lipid oxidation products are TAG with a hydroperoxy, hydroxy, epoxy, or oxo (ketone or aldehyde) group or combination of these groups. The breakdown of TAG hydroperoxides yields nonvolatile glyceride species with two intact fatty acid chains and one short chain mostly ending in an aldehyde or hydroxy group (2 1/2 glycerides). By means of normal-phase high-performance liquid chromatography (HPLC) with mass spectrometric (MS) detection, non-volatile lipid oxidation products can be separated according to polarity. This results in separation into classes of TAG oxidation products, such as epoxy-TAG, oxo-TAG, hydroperoxy-TAG, hydroxy-TAG and 2 1/2 glycerides, which can be identified using selected ion chromatograms. The retention times of TAG oxidation products on the normal-phase HPLC system and the signal intensity of the MS detector are stable enough to enable quantitative analysis based on external calibration. The normal-phase HPLC-MS method is very suitable for the characterization and quantitation of nonvolatile TAG oxidation products in oxidized TAG reference compounds as well as in real oils or oil phases isolated from emulsions, spreads, or other fat-based food products. This method can give detailed information for the study of lipid oxidation mechanisms.

Analysis of Non-volatile Constituents in Dracocephalum Species by HPLC and GC-MS

Abstract  In this paper we identify/determine the composition of the extracts of Dracocephalum moldavica L. and D. ruyschiana L. with special emphasis on their flavonoids, aliphatic, aromatic carboxylic acids and sugars. The plant materials were extracted using methanol-acetone 1:1 (v/v) acidified with HCl (0.05%). Composition of extract's most red fractions was identified by HPLC, while the constituents of the entire extract were identified and quantified as their trimethylsilyl-(oxime) ether/ester derivatives by GC-MS. On the basis of HPLC analyses - (performed on Phenomenex Luna 5 μm C18 column, 250× 4.6 mm I.D.; eluent: acetonitrile - trifluoroacetic acid (0.1 %); isocratic/gradient conditions) - delphinidin, cyanidin and pelargonidin (in traces) were identified by spectral analysis and on the basis of authentic standard's addition. GC-MS analyses were carried out immediately with extracts, as well as subsequently to extract's hydrolysis (trifluoroacetic acid: 2M, 2h, 4h). Constituents were identified and quantified as their trimethylsilyl-(oxime) ether/ester derivatives, from a single run, on the basis of their total (TIC) and selective fragment ion (SIM) responses. Calculations were related to the dry matter content of extracts. As main constituents monosaccharides, sugar alcohols di- and trisaccharides, aliphatic (phosphoric, succinic, levulinic, malic, tartaric, fatty acids) and aromatic (quinic, chlorogenic, caffeic, ferulic, 3,4-dihydroxyphenyllactic, rosmarinic) carboxylic acids and their corresponding esters; apigenin, luteolin flavon aglycons and tocopherol, in total 33 constituents were quantitated partly by their TIC, partly by their SIM responses. Identification/quantification proved to be in total of 35–69% (expressed in the dry matter content of extracts).

Analysis of mechanisms underlying BRMS1 suppression of metastasis

Abstract  Introduction of normal, neomycin-tagged human chromosome 11 (neo11) reduces the metastatic capacity of MDA-MB-435 human breast carcinoma cells by 70–90% without affecting tumorigenicity. Differential display comparing MDA-MB-435 and neo11/435 led to the discovery of a human breast carcinoma metastasis suppressor gene, BRMS1, which maps to chromosome 11q13.1–q13.2. Stable transfectants of MDA-MB-435 and MDA-MB-231 breast carcinoma cells with BRMS1 cDNA still form progressively growing, locally invasive tumors when injected in mammary fat pads of athymic mice but exhibit significantly lower metastatic potential (50–90% inhibition) to lungs and regional lymph nodes. To begin elucidating the mechanism(s) of action, we measured the ability of BRMS1 to perturb individual steps of the metastatic cascade modeled in vitro. Consistent differences were not observed for adhesion to extracellular matrix components (laminin, fibronectin, type IV collagen, type I collagen, Matrigel); growth rates in vitro or in vivo; expression of matrix metalloproteinases, heparanase, or invasion. Likewise, BRMS1 expression did not up regulate expression of other metastasis suppressors, such as NM23, Kai1, KiSS1 or E-cadherin. Motility of BRMS1 transfectants was modestly inhibited (30–60%) compared to parental and vector-only transfectants. Ability to grow in soft agar was also decreased in MDA-MB-435 cells by 80–89%, but the decrease for MDA-MB-231 was less (13–15% reduction). Also, transfection and re-expression of BRMS1 restored the ability of human breast carcinoma cells to form functional homotypic gap junctions. Collectively, these data suggest that BRMS1 suppresses metastasis of human breast carcinoma by complex, atypical mechanisms.

Analysis of manure and soil nitrogen mineralization during incubation

Abstract  Understanding the N-cycling processes that ensue after manuring soil is essential in order to estimate the value of manure as an N fertilizer. A laboratory incubation of manured soil was carried out in order to study N mineralization, gas fluxes, denitrification, and microbial N immobilization after manure application. Four different manures were enclosed in mesh bags to allow for the separate analysis of manure and soil. The soils received 0.15 mg manure N g–1 soil, and the microcosms were incubated aerobically and sampled throughout a 10-week period. Manure addition resulted in initial NH4-N concentrations of 22.1 to 36.6 mg kg–1 in the microcosms. All manured microcosms had net declines in soil mineral N. Denitrification resulted in the loss of 14.7 to 39.2% of the added manure N, and the largest N losses occurred in manures with high NH4-N content. Increased soil microbial biomass N amounted to 6.0 to 8.6% of the added manure N. While the microcosms as a whole had negative N mineralization, all microcosms had positive net nitrification within the manure bags. Gas fluxes of N2O and CO2 increased in all manured soils relative to the controls. Our results show that measurement of microbial biomass N and denitrification is important to understand the fate of manure N upon soil application.

Analysis of long-chain ω-3 fatty acid content in fish-oil supplements

Zusammenfassung  HINTERGRUND: In zahlreichen Studien wurden die verschiedensten vorteilhaften Effekte von mehrfach unges?ttigten Omega 3 Fetts?uren auf Atherosklerose, Arrhythmie und Hypertriglyzerid?mie nachgewiesen, was zahlreiche Gesundheitsorganisationen dazu veranlasst hat, einen t?glichen Verzehr von einem Gramm Omega 3 Fetts?uren t?glich für antiatherosklerotische sowie antiarrhytmische Wirkungen oder zwei bis vier g/d Omega 3 Fetts?uren zur Senkung der Plasmatriglyzeride zu empfehlen. Es sind zahlreiche Pr?parate auf dem Markt erschienen, welche die ω-3 PUFA-arme westliche Ern?hrung in Form von Kapseln erg?nzen. Da diese Pr?parate betr?chtlichen Variationen des Gehalts an langkettigen ω-3 PUFAs unterworfen sein k?nnen, haben wir neun kommerziell erh?ltliche Produkte bezugnehmend auf ihre Fetts?urekomposition getestet. METHODEN: Neun kommerziell erh?ltliche ω-3 PUFA Nahrungserg?nzungsmittel wurden mittels kapill?rer Gaschromatographie auf ihren Gehalt an langkettigen ω-3 PUFA untersucht. ERGEBNISSE: Die neun von uns getesteten Pr?parate zeigen hinsichtlich der Konzentration an langkettigen ω-3 PUFA gro?e Unterschiede von bis zu 63,7 ± 1,58 mol % (p = 0,002) und die Meisten scheitern daran, die empfohlene Tagesdosis von einem Gramm zu erzielen, selbst wenn sie in der h?chsten vom Hersteller angegebenen Dosierung verabreicht werden. Acht der Pr?parate enthalten entweder gleiche oder signifikant h?here langkettige ω-3 PUFA Mengen als vom Hersteller angegeben und ein Hersteller macht keine Angabe. Die h?chsten Anteile an Eicosapentaens?ure (EPA) und Docosahexaens?ure (DHA) wurden in Omacor? (95,80 ± 0,63%) und Percucor? (76,8 ± 7,11%) vorgefunden. KONKLUSION: Verabreichung von langkettigen ω-3 Fetts?urepr?paraten kann in gro?en Unterschieden der tats?chlich konsumierten Menge resultieren. Daher ist es empfehlenswert, die am h?chsten standardisierten und gereinigten Produkte zu verwenden.

Analysis of lipoxygenase-derived fatty acid hydroperoxides by electrospray ionization tandem mass spectrometry

Abstract  A rapid method is described for the identification of fatty acid hydroperoxides by electrospray ionization-tandem mass spectrometry coupled to liquid chromatography without any derivatization required prior to analysis. Localization of fatty acid hydroperoxides in complex mixtures was achieved by monitoring the loss of hydrogen peroxide using constant neutral loss scanning. In the presence of 5 mM NH4OAc in methanol-water, adductions [M+NH4]+ were formed almost exclusively, directly revealing the molecular mass of the thermolabile hydroperoxides. In addition, low energy collision-induced dissociation of precursor ions [M+NH4]+ led to characteristic product ions from both the 9- and 13-regioisomers. Thus, electrospray ionization-tandem mass spectrometry provides a straightforward approach for the study of the regioselectivity of lipoxygenase catalysis without any derivatization step required prior to analysis.

Analysis of isomeric diacylglycerolic classes to evaluate the quality of olive oil in relation to storage conditions

Abstract  The % content and the % fatty acid (FA) composition of the three isomeric classes of diacylglycerol (DAG) fraction (sn-1,2-, sn-2,3-, sn-1,3-DAG) were valued in four groups of extra virgin olive oils, stored in different conditions of temperature and time, with the objective to study the influence of the cited parameters on olive oil DAG classes. The separation and % quantitation of all isomeric DAG classes were carried out by normal phase-high performance liquid chromatography (NP-HPLC) on total DAG fraction previously isolated by TLC and subjected to derivatization step with (S)-(+)-(1-naphtyl)-ethylisocyanate; moreover the % FA composition of each DAG class was determined by HRGC analysis. The results showed notable differences on the % content of each DAG class as well as the ratios between the classes. The samples analysed as soon as they were produced showed the highest % contents for sn-1,2-DAG and the lowest for sn-1,3- and sn-2,3-DAG. The samples stored at 30 °C generally showed the highest % contents of sn-1,3-DAG, because of the isomerization processes of sn-1,2-DAG to the more stable sn-1,3-DAG. The results of structural analysis of the sn-1,2-DAG class besides confirmed that isomerization processes occurred and that the storage temperature had a predominant role; this research emphasizes the importance of the valuation of % contents together with the structural analysis of the isomeric DAG classes as useful analytical parameters to evaluate the storage conditions and the conservation status of olive oils.

Analysis of interaction partners for perilipin and ADRP on lipid droplets?

Abstract  Despite the critical roles of intracellular lipid droplets (LDs) in lipid storage and metabolism, little is known about the molecular mechanisms of their functions. Several protein components associated with the surface of LDs have been identified. A major one is perilipin in adipocytes and steroidogenic cells, whereas ADRP in most other cell types. They are loosely grouped as a small protein family sharing a common N-terminal motif, called the PAT domain. Perilipin regulates the breakdown of triacylglycerol in LDs via its phosphorylation. ADRP is characterized as a fatty acid binding protein and involved in lipid uptake and LD formation. For examining the functions of perilipin and ADRP at the molecular level, we performed yeast two-hybrid screening in this study, to find their functional partners. We identified CGI-58, a product of the causal gene of Chanarin-Dorfman syndrome (CDS), as an interactor for both perilipin and ADRP. Specific interaction between CGI-58 and perilipin was confirmed in a GST-pulldown assay and supported by fluorescence microscopic analyses. We further demonstrated that CGI-58 is principally located at the surface of LDs in 3T3-L1 cells, together with perilipin, and its expression is upregulated upon stimulation for adipocyte differentiation. Other than CGI-58, we also identified in yeast two-hybrid screening HSP86 and D52 tumor proteins as binding partners of perilipin, and IRG-47 of ADRP. These factors might be cooperated with perilipin and ADRP, and hence involved in membrane dynamics of LDs as well as the regulation of lipolysis on the surface of LDs.

Analysis of Human Oral Mucosa Ex Vivo for Fatty Acid Compositions Using Fourier-transform Infrared Spectroscopy

Abstract  The purpose of this research was to develop an infrared spectroscopic technique (Fourier transform infrared spectroscopy with attenuated total reflectance system; FTIR-ATR) for non-invasive measurement of saturated and unsaturated fatty acid compositions in human oral mucosa obtained from three nationalities; Iranian, Vietnamese, and Indonesian. The histogram patterns of fatty acid compositions for three nationalities suggest that the pattern of unsaturated fatty acids were quite different, although the distribution profiles of fatty acid to lipid ratios in FTIR-ATR has a similar normal pattern with small difference in skewness and mode. The second derivative infrared spectra of the mucosal tissues in the wavenumber regions from 1,600 to 1,760 cm−1 and 2,800 to 3,050 cm−1 were analyzed with partial least squares (PLS) multivariate regression analysis method. With this analysis method we compared predicted values with the measured values of ten categorized fatty acid compositions, i.e., a(saturated C17 or lower), b(C16:1 + C17:1), c(C18:0), d(C18:1), e(C18:2), f(saturated C20 or longer), g(C20:3 + C20:4), h(C22:1 + C24:1), i(C22:6), j(γC18:3). Almost all fatty acid compositions of oral mucosa were well predicted with differences between predicted and measured values within ±5% of total, however, errors were relatively larger in minor components such as C22:6 than major components.

Analysis of genetic diversity and phylogenetic relationships among red jungle fowls and Chinese domestic fowls

Abstract  Genetic diversity and phylogenetic relationships among 568 individuals of two red jungle fowl subspecies (Gallus gallus spadiceus in China and Gallus gallus gallus in Thailand) and 14 Chinese domestic chicken breeds were evaluated with 29 microstaellite loci, the genetic variability within population and genetic differentiation among population were estimated, and then genetic diversity and phylogenetic relationships were analyzed among red jungle fowls and Chinese domestic fowls. A total of 286 alleles were detected in 16 population with 29 microsatellite markers and the average number of the alleles observed in 29 microsatellite loci was 9.86±6.36. The overall expected heterozygosity of all population was 0.6708±0.0251, and the number of population deviated from Hardy-Weinberg equilibrium per locus ranged from 0 to 7. In the whole population, the average of genetic differentiation among population, measured as FST value, was 16.7% (P<0.001), and all loci contributed significantly (P<0.001) to this differentiation. It can also be seen that the deficit of heterozygotes was very high (0.015) (P<0.01). Reynolds’ distance values varied between 0.036 (Xiaoshan chicken-Luyuan chicken pair) and 0.330 (G. gallus gallus-Gushi chicken pair). The Nm value ranged from 0.533 (between G. gallus gallus and Gushi chicken) to 5.833 (between Xiaoshan chicken and Luyuan chicken). An unrooted consensus tree was constructed using the neighbour-joining method and the Reynolds’ genetic distance. The heavy-body sized chicken breeds, Luyuan chicken, Xiaoshan chicken, Beijing Fatty chicken, Henan Game chicken, Huainan Partridge and Langshan chicken formed one branch, and it had a close genetic relationship between Xiaoshan chicken-Luyuan chicken pair and Chahua chicken-Tibetan chicken pair. Chahua chicken and Tibetan chicken had closer genetic relationship with these two subspecies of red jungle fowl than other domestic chicken breeds. G. gallus spadiceus showed closer phylogenetic relationship with Chinese domestic chicken breeds than G. gallus gallus. All 29 microstaellite loci in this study showed high levels of polymorphism and significant genetic differentiation was observed among two subspecies of red jungle fowl and 14 Chinese domestic chicken breeds. The evolutional dendrogram is as follows: evolutional breeds→primitive breeds (Chahua chicken and Tibetan)→red jungle fowl in China (G. gallus spadiceus)→red jungle fowl in Thailand (G. gallus gallus). The results supported the theory that the domestic fowls might originate from different subspecies of red jungle fowl and Chinese domestic fowls had independent origin.

Analysis of ethoxylated fatty amines. Comparison of methods for the determination of molecular weight

Abstract  Specific lengths of the fatty and polyoxyethylene chains of ethoxylated fatty amines are critical to their performance in specific applications, and thus the ability to characterize these surfactants accurately is crucial. Normal-phase high-performance liquid chromatography (HPLC) and matrixassisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry methods were developed to determine with accuracy the molecular weight and degree of ethoxylation of ethoxylated fatty amines. Ethoxylated fatty amines were analyzed using these methods, and comparison was made to molecular weight determinations using proton nuclear magnetic resonance (NMR), neutralization equivalent weight, and hydroxyl value methods. Molecular weight results from normalphase HPLC analyses were in very good agreement with MALDI-TOF results, typically varying less than one ethylene oxide unit. A reversed-phase HPLC method was developed to determine concentrations of polyethylene glycols (PEG) and fatty homologs. PEG interfered with molecular weight determinations by NMR, neutralization equivalent weight, and hydroxyl value methods. PEG caused no interference with molecular weight determinations by normal-phase HPLC and MALDI-TOF methods.

Analysis of endophytic bacterial communities of potato by plating and denaturing gradient gel electrophoresis (DGGE) of 16S rDNA based PCR fragments

Abstract  The diversity of endophytic bacterial populations of potato (Solanum tuberosum cv Desirée) was assessed using a combination of dilution plating of plant macerates followed by isolation and characterization of isolates, and direct PCR-DGGE on the basis of DNA extracted from plants. The culturable endophytic bacterial communities detected in potato stem bases as well as in roots were in most cases on the order 103 to 105 CFU g−1 of fresh plant tissue. Dilution plating revealed that a range of bacterial types dominated these populations. Dominant isolates fell into the α and γ subgroups of the Proteobacteria, as well as in the Flavobacterium/Cytophaga group. Different representatives of the Firmicutes were also found. The most frequently isolated strains (>5% of the total) were characterized as different Pseudomonas spp. (including P. aureofaciens, P. corrugata, and P. putida), Agrobacterium radiobacter, Stenotrophomonas maltophilia, and Flavobacterium resinovorans, using fatty acid methyl ester (FAME) analysis and/or sequencing of their partial 16S ribosomal RNA genes. Other Proteobacteria or Firmicutes were also found, albeit infrequently, and mainly in potato stem tissue. The fate of three putative potato endophytes, Stenotrophomonas maltophilia, Bacillus sp., and Sphingomonas paucimobilis, was monitored following their release into potato plants via injection, via root dipping, or via the soil. Following stem injection, the S. maltophilia and Bacillus inoculants could be tracked over time periods of, respectively, 22 and 1 day(s) by dilution plating as well as via PCR-DGGE. However, only S. maltophilia was able to colonize, and persist in, plant tissue from soil or dipped roots. S. paucimobilis was never recovered from the plant irrespective of the mode of introduction. The diversity of the indigenous bacterial flora associated with potato was then monitored via PCR-DGGE. The patterns obtained revealed the existence of bacterial communities of limited complexity, with communities from potato stems typically differing from those from stem peel and roots. Evidence was obtained for the endophytic occurrence of a range of organisms falling into the α, β, and γ subgroups of the Proteobacteria as well as in the Firmicutes. Several of the sequences found matched those from isolates, suggesting that the molecular evidence reported culturable organisms. However, a number of sequences did not have matching sequences from isolates, suggesting that non-culturable or as-yet-uncultured endophytic organisms were being detected.

Analysis of embryo, cytoplasm and maternal effects on fatty acid components in soybean ( Glycine max Merill.)

Abstract  The quality of oil determined by the constituents and proportion of fatty acid components, and the understanding of heredity of fatty acid components are of importance to breeding good quality soybean varieties. Embryo, cytoplasmic and maternal effects and genotype × environment interaction effects for quality traits of soybean [Glycine max (L.) Merrill.] seeds were analyzed using a general genetic model for quantitative traits of seeds with parents, F1 and F2, of 20 crosses from a diallel mating design of five parents planted in the field in 2003 and 2004 in Harbin, China. The interaction effects of palmitic, stearic, and linoleic acid contents were larger than the genetic main effects, while the genetic main effects were equal to interaction effects for linolenic and oleic acid content. Among all kinds of genetic main effects, the embryo effects were the largest for palmitic, stearic, and linoleic acids, while the cytoplasm effects were the largest for oleic and linolenic acids. Among all kinds of interaction effects, the embryo interaction effects were the largest for fatty acids. The sum of additive and additive × environment effects were larger than that of dominance and dominance × environment effects for the linolenic acid content, but not for other quality traits. The general heritabilities were the main parts of heritabilities for palmitic and oleic acid contents, but the interaction was more important for stearic, linoleic, and linolenic acid contents. For the general heritability, maternal and cytoplasm heritabilities were the main components for palmitic, oleic, and linolenic acid contents. It was shown for the interaction heritabilities that the embryo interaction heritabilities were more important for oleic and linolenic acid contents, while the maternal interaction heritabilities were more important for linoleic acid content. Among selection response components, the maternal and cytoplasm general responses and/or interaction responses were more important for palmitic, stearic, oleic, and linoleic acid contents. The main selection response components were from the embryo general response and/or interaction response for linolenic acid content. It suggested that the selection of palmitic, stearic, oleic, and linoleic acid contents in offspring should be in maternal plants, while linolenic acid content should be improved by screening or selecting the single seed in higher generations.

Analysis of diurnal and vertical microbial diversity of a hypersaline microbial mat

Abstract  Microbial mats are prokaryotic communities that provide model systems to analyze microbial diversity and ecophysiological interactions. Community diversity of microbial mat samples was assessed at 8:00 a.m. and 3:00 p.m. in a combined analysis consisting of 16S rRNA-denaturing gradient gel electrophoresis (DGGE) and phospholipid fatty acid (PLFA) profiles. The divergence index determined from PLFA and DGGE data showed that depth-related differences have a greater influence on diversity than temporal variations. Shannon and Simpson indices yielded similar values in all samples, which suggested the stable maintenance of a structurally diverse microbial community. The increased diversity observed at 3:00 p.m. between 2.5 and 4 mm can be explained mainly by diversification of anaerobic microorganisms, especially sulfate-reducing bacteria. In the afternoon sampling, the diversity index reflected a higher diversity between 4 and 5.5 mm depth, which suggested an increase in the diversity of strict anaerobes and fermenters. The results are consistent with the conclusion that hypersaline microbial mats are characterized by high degree of diversity that shifts in response to the photobiological adaptations and metabolic status of the microbial community.