Synthesis of triheptanoin and formulation as a solid diet for rodents

Triheptanoin-enriched diets have been successfully used in the experimental treatment of various metabolic disorders. Maximal therapeutic effect is achieved in the context of a ketogenic diet where triheptanoin oil provides 3040% of the daily caloric intake. However, pre-clinical studies using triheptanoin-rich diets are hindered by the difficulty of administering to laboratory animals as a solid foodstuff. In the present study, we successfully synthesized triheptanoin to the highest standards of purity from glycerol and heptanoic acid, using sulfonated charcoal as a catalyst. Triheptanoin oil was then formulated as a solid, stable and palatable preparation using a ketogenic base and a combination of four commercially available formulation agents: hydrophilic fumed silica, hydrophobic fumed silica, microcrystalline cellulose, and talc. Diet compliance and safety was tested on C57Bl/6 mice over a 15-week period, comparing overall status and body weight change. Practical applications: This work provides a complete description of (i) an efficient and cost-effective synthesis of triheptanoin and (ii) its formulation as a solid, stable, and palatable ketogenic diet (triheptanoin-rich; 39% of the caloric intake) for rodents. Triheptanoin-rich diets will be helpful on pre-clinical experiments testing the therapeutic efficacy of triheptanoin in different rodent models of human diseases. In addition, using the same solidification procedure, other oils could be incorporated into rodent ketogenic diet to study their dosage and long-term effects on mammal health and development. This approach could be extremely valuable as ketogenic diet is widely used clinically for epilepsy treatment.

Postmortem angiography after vascular perfusion with diesel oil and a lipophilic contrast agent.

OBJECTIVE: The objective of our study was to establish optimal perfusion conditions for high-resolution postmortem angiography that would permit dynamic visualization of the arterial and venous systems. MATERIALS AND METHODS: Cadavers of two dogs and one cat were perfused with diesel oil through a peristaltic pump. The lipophilic contrast agent Lipiodol Ultra Fluide was then injected, and angiography was performed. The efficiency of perfusion was evaluated in the chick chorioallantoic membrane. RESULTS: Vessels could be seen up to the level of the smaller supplying and draining vessels. Hence, both the arterial and the venous sides of the vascular system could be distinguished. The chorioallantoic membrane assay revealed that diesel oil enters microvessels up to 50 microm in diameter and that it does not penetrate the capillary network. CONCLUSION: After establishing a postmortem circulation by diesel oil perfusion, angiography can be performed by injection of Lipiodol Ultra Fluide. The resolution of the images obtained up to 3 days after death is comparable to that achieved in clinical angiography.

Flight, feeding and reproductive behavior of Phyllophaga cuyabana (Moser) (Coleoptera: Melolonthidae) adults

Phyllophaga cuyabana is a univoltine species and its development occurs completely underground. Its control by conventional methods, such as chemical and biological insecticides, is difficult, so it is important to understand its dispersion, reproduction, and population behavior in order to determine best pest management strategies. The objective of this work was to study the behavior of adults of P. cuyabana. This study was carried out in the laboratory, greenhouse and field sites in Paraná State, Brazil (24º25' S and 52º48' W), during four seasons. The results obtained demonstrate that: a) P. cuyabana adults have a synchronized short-flight period when mating and reproduction occurs; b) adults tend to aggregate in specific sites for mating; c) the majority of adults left the soil on alternate nights; d) the choice of mating and oviposition sites was made by females before copulation, since after copulation adults did not fly from or bury themselves at nearby locations; e) females that fed on leaves after mating, oviposited more eggs than females that had not fed;f) plant species such as sunflower (Helianthus annuus) and the Crotalaria juncea are important food sources for adults.

A multimodality vascular imaging phantom with fiducial markers visible in DSA, CTA, MRA, and ultrasound.

The objective was to design a vascular phantom compatible with digital subtraction angiography, computerized tomography angiography, ultrasound and magnetic resonance angiography (MRA). Fiducial markers were implanted at precise known locations in the phantom to facilitate identification and orientation of plane views from three-dimensional (3-D) reconstructed images. A vascular conduit connected to tubing at the extremities of the phantom ran through an agar-based gel filling it. A vessel wall in latex was included around the conduit to avoid diffusion of contrast agents. Using a lost-material casting technique based on a low melting point metal, geometries of pathological vessels were modeled. During the experimental testing, fiducial markers were detectable in all modalities without distortion. No leak of gadolinium through the vascular wall was observed on MRA after 5 hours. Moreover, no significant deformation of the vascular conduit was noted during the fabrication process (confirmed by microtome slicing along the vessel). The potential use of the phantom for calibration, rescaling, and fusion of 3-D images obtained from the different modalities as well as its use for the evaluation of intra- and inter-modality comparative studies of imaging systems are discussed. In conclusion, the vascular phantom can allow accurate calibration of radiological imaging devices based on x-ray, magnetic resonance and ultrasound and quantitative comparisons of the geometric accuracy of the vessel lumen obtained with each of these methods on a given well defined 3-D geometry.

Use of solaria to predict weed density and floristic composition in no-till cropping systems

The objective of this work was to evaluate the efficiency of a new method, developed for predicting density and floristic composition of weed communities in field crops. Based on the use of solaria (100 mm transparent plastic tarps lying on the soil) to stimulate weed seedlings emergence, the method was tested in Tandil, Argentina, from 1998 to 2001. The system involved corn and sunflower in commercial no-till system. Major weeds in the experiments included Digitaria sanguinalis, Setaria verticillata and S. viridis, which accounted for 98% of the weed community in the three years of experiments since 1998. Large numbers of Tagetes minuta, Chenopodium album and Ammi majus were present in 2001. Comparison of weed communities under solaria with communities in field crops indicated that the method is useful for predicting the presence and density of some major weed species, at both high and low densities, of individuals in areas of 10 ha using only five solaria. Low density of weed species makes the method particularly useful to help deciding the time for herbicide applications to avoid soil contamination.

Direct evidence for the existence of dairying farms in prehistoric Central Europe (4th millennium BC)

The molecular and isotopic chemistry of organic residues from archaeological potsherds was used to obtain further insight into the dietary trends and economies at the Constance lake-shore Neolithic settlements. The archaeological organic residues from the Early Late Neolithic (3922-3902 BC) site Hornstaad-Hornle IA/Germany are, at present, the oldest archaeological samples analysed at the Institute of Mineralogy and Geochemistry of the University of Lausanne. The approach includes 13C/12C and 15N/14N ratios of the bulk organic residues, fatty acids distribution and 13C/12C ratios of individual fatty acids. The results are compared with those obtained from the over 500 years younger Neolithic (3384-3370 BC) settlement of Arbon Bleiche 3/Switzerland and with samples of modern vegetable oils and fat of animals that have been fed exclusively on C3 forage grasses. The overall fatty acid composition (C9 to C24 range, maximizing at C14 and C16), the bulk 13C/12C and 15N/14N ratios (delta13C, delta15N) and the 13C/12C ratios of palmitic (C16:0), stearic (C18:0) and oleic acids (C18:1) of the organic residues indicate that most of the studied samples (25 from 47 samples and 5 from 41 in the delta13C18:0 vs. delta13C16:0 and delta13C18:0 vs. delta13C18:1 diagrams, respectively) from Hornstaad-Hornle IA and Arbon Bleiche 3 sherds contain fat residues of pre-industrial ruminant milk, and young suckling calf/lamb adipose. These data provide direct proof of milk and meat (mainly from young suckling calves) consumption and farming practices for a sustainable dairying in Neolithic villages in central Europe around 4000 BC.dagger

Development of Non-Petroleum Based Binders for Use in Flexible Pavements, TR-594, 2010

Most bituminous adhesives or binders that are used for pavement materials are derived primarily from fossil fuels. With petroleum oil reserves becoming depleted and the drive to establish a bio-based economy, there is a push to produce binders from alternative sources, particularly from biorenewable resources. However, until now, no research has studied the applicability of utilizing bio-oils as a bitumen replacement (100% replacement) in the pavement industry. The main objective of this research was to test various properties of bio-oils in order to determine the applicability of using bio-oils as binders in the pavement industry. The overall conclusions about the applicability of using bio-oils as bio-binders in the pavement industry can be summarized as follows: 1. Bio-oils cannot be used as bio-binders/pavement materials without any heat pre-treatment/upgrading procedure. 2. Current testing standards and specifications, especially Superpave procedures, should be modified to comply with the properties of bio-binders. 3. The temperature range of the viscous behavior for bio-oils may be lower than that of bitumen binders by about 30°–40° C. 4. The rheological properties of the unmodified bio-binders vary in comparison to bitumen binders, but the rheological properties of these modified bio-binders change significantly upon adding polymer modifiers. 5. The high-temperature performance grade for the developed bio-binders may not vary significantly from that of the bitumen binders, but the low-temperature performance grade may vary significantly

Blue light activates a specific protein kinase in higher plants.

Blue light mediates the phosphorylation of a membrane protein in seedlings from several plant species. When crude microsomal membrane proteins from dark-grown pea (Pisum sativum L.), sunflower (Helianthus annuus L.), zucchini (Cucurbita pepo L.), Arabidopsis (Arabidopsis thaliana L.), or tomato (Lycopersicon esculentum L.) stem segments, or from maize (Zea mays L.), barley (Hordeum vulgare L.), oat (Avena sativa L.), wheat (Triticum aestivum L.), or sorghum (Sorghum bicolor L.) coleoptiles are illuminated and incubated in vitro with [gamma-(32)P]ATP, a protein of apparent molecular mass from 114 to 130 kD is rapidly phosphorylated. Hence, this system is probably ubiquitous in higher plants. Solubilized maize membranes exposed to blue light and added to unirradiated solubilized maize membranes show a higher level of phosphorylation of the light-affected protein than irradiated membrane proteins alone, suggesting that an unirradiated substrate is phosphorylated by a light-activated kinase. This finding is further demonstrated with membrane proteins from two different species, where the phosphorylated proteins are of different sizes and, hence, unambiguously distinguishable on gel electrophoresis. When solubilized membrane proteins from one species are irradiated and added to unirradiated membrane proteins from another species, the unirradiated protein becomes phosphorylated. These experiments indicate that the irradiated fraction can store the light signal for subsequent phosphorylation in the dark. They also support the hypothesis that light activates a specific kinase and that the systems share a close functional homology among different higher plants.

Combined effects of endurance training and dietary unsaturated fatty acids on physical performance, fat oxidation and insulin sensitivity.

Endurance training improves exercise performance and insulin sensitivity, and these effects may be in part mediated by an enhanced fat oxidation. Since n-3 and n-9 unsaturated fatty acids may also increase fat oxidation, we hypothesised that a diet enriched in these fatty acids may enhance the effects of endurance training on exercise performance, insulin sensitivity and fat oxidation. To assess this hypothesis, sixteen normal-weight sedentary male subjects were randomly assigned to an isoenergetic diet enriched with fish and olive oils (unsaturated fatty acid group (UFA): 52 % carbohydrates, 34 % fat (12 % SFA, 12 % MUFA, 5 % PUFA), 14 % protein), or a control diet (control group (CON): 62 % carbohydrates, 24 % fat (12 % SFA, 6 % MUFA, 2 % PUFA), 14 % protein) and underwent a 10 d gradual endurance training protocol. Exercise performance was evaluated by measuring VO2max and the time to exhaustion during a cycling exercise at 80 % VO2max; glucose homeostasis was assessed after ingestion of a test meal. Fat oxidation was assessed by indirect calorimetry at rest and during an exercise at 50 % VO2max. Training significantly increased time to exhaustion, but not VO2max, and lowered incremental insulin area under the curve after the test meal, indicating improved insulin sensitivity. Those effects were, however, of similar magnitude in UFA and CON. Fat oxidation tended to increase in UFA, but not in CON. This difference was, however, not significant. It is concluded that a diet enriched with fish- and olive oil does not substantially enhance the effects of a short-term endurance training protocol in healthy young subjects.

Lipase Improvement: Goals and strategies

Lipases have received great attention as industrial biocatalysts in areas like oils and fats processing, detergents, baking, cheese making, surface cleaning, or fine chemistry . They can catalyse reactions of insoluble substrates at the lipid-water interface, preserving their catalytic activity in organic solvents. This makes of lipases powerful tools for catalysing not only hydrolysis, but also various reverse reactions such as esterification, transesterification, aminolysis, or thiotransesterifications in anhydrous organic solvents. Moreover, lipases catalyse reactions with high specificity, regio and enantioselectivity, becoming the most used enzymes in synthetic organic chemistry. Therefore, they display important advantages over classical catalysts, as they can catalyse reactions with reduced side products, lowered waste treatment costs, and under mild temperature and pressure conditions. Accordingly, the use of lipases holds a great promise for green and economical process chemistry.

Dietary n-6- or n-3-rich vegetable fats and antioxidants: effects on fatty acid composition and stability of rabbit plasma, liver and meat

We supplemented diets with a-tocopheryl acetate (100 mg/kg) and replaced beef tallow (BT) in feeds with increasing doses of n-6- or n-3-rich vegetable fat sources (linseed and sunflower oil), and studied the effects on the fatty acid (FA) composition, the a-tocopherol (aT) content and the oxidative stability of rabbit plasma and liver. These effects were compared with those observed in a previous study in rabbit meat. As in meat, the content of saturated, monounsaturated and trans FA in plasma and liver mainly reflected feed FA profile, except stearic acid in liver, which increased as feeds contained higher doses of vegetable fat, which could be related to an inhibition of the activity of the stearoyl-CoA-desaturase. As linseed oil increased in feeds, the n-6/n-3 FA ratio was decreased in plasma and liver as a result of the incorporation of FA from diets and also, due to the different performance and selectivity of desaturase enzymes. However, an increase in the dose of vegetable fat in feeds led to a significant reduction in the aT content of plasma and liver, which was greater when the fat source was linseed oil. Increasing the dose of vegetable fat in feeds also led to an increase in the susceptibility to oxidation (lipid hydroperoxide (LHP) value) of rabbit plasma, liver and meat and on the thiobarbituric acid (TBA) values of meat. Although the dietary supplementation with a-tocopheryl acetate increased the aT content in plasma and liver, it did not modify significantly their TBA or LHP values. In meat however, both TBA and LHP values were reduced by the dietary supplementation with a-tocopheryl acetate. The plasma aT content reflected the aT content in tissues, and correlated negatively with tissue oxidability. From the studied diets, those containing 1.5% linseed oil plus 1.5% BT and 100 mg of a-tocopheryl acetate/kg most improved the FA composition and the oxidative stability of rabbit tissues.

Cyclosporine A delivery to the eye: a pharmaceutical challenge.

Systemic administration of cyclosporine A (CsA) is commonly used in the treatment of local ophthalmic conditions involving cytokines, such as corneal graft rejection, autoimmune uveitis and dry eye syndrome. Local administration is expected to avoid the various side effects associated with systemic delivery. However, the currently available systems using oils to deliver CsA topically are poorly tolerated and provide a low bioavailability. These difficulties may be overcome through formulations aimed at improving CsA water solubility (e.g. cyclodextrins), or those designed to facilitate tissue drug penetration using penetration enhancers. The use of colloidal carriers (micelles, emulsions, liposomes and nanoparticles) as well as the approach using hydrosoluble prodrugs of CsA have shown promising results. Solid devices such as shields and particles of collagen have been investigated to enhance retention time on the eye surface. Some of these topical formulations have shown efficacy in the treatment of extraocular diseases but were inefficient at reaching intraocular targets. Microspheres, implants and liposomes have been developed to be directly administered subconjunctivally or intravitreally in order to enhance CsA concentration in the vitreous. Although progress has been made, there is still room for improvement in CsA ocular application, as none of these formulations is ideal.

Fish oil supplementation does not alter energy efficiency in healthy males

BACKGROUND AND AIMS: Fish oil (FO) supplementation prevents the development of obesity and insulin resistance, and upregulate the expression of UCP3 in skeletal muscle in rodents. This may represent indirect evidence that FO promotes fat oxidation and/or alter energy efficiency. The aim of this study was to evaluate whether such effects can be observed in humans. The metabolic effects of FO were assessed during exercise in order to obtain a direct measurement of energy efficiency. METHODS: Eight healthy male volunteers were studied with and without supplementation with 7.2 g/day FO (including 1.1 g/day eicosopentaenoic acid and 0.7 g/day decosahexaenoic acid) during 14 days. Their VO(2 max) was measured on cycle ergometer. Thereafter, energy metabolism (substrate oxidation, energy expenditure and energy efficiency) was assessed during a 30 min cycling exercise at 50% VO(2 max) performed 2 h 30 after a standardized, high carbohydrate breakfast. RESULTS: VO(2 max) was 38.6+/-2.2 after FO and 38.4+/-2.0 (mL x kg(-1) x min(-1)) in control conditions (NS). Basal plasma glucose, insulin and NEFA concentrations, and energy metabolism were similar with FO and in controls. During exercise, the increases in plasma NEFA concentrations, energy expenditure, glucose and lipid oxidation, and the decreases in glycaemia and insulinemia were not altered by FO intake. Energy efficiency was 22.4+/-0.6% after FO vs 21.8+/-0.7% in controls. In order to ascertain that the absence of effects of FO was not due to consumption of a carbohydrate meal immediately before exercise, 4 of the 8 subjects were re-studied in fasting conditions, FO also failed to alter energy efficiency in this subset of studies. CONCLUSION: FO supplementation did not significantly alter energy metabolism and energy efficiency during exercise in healthy humans.

Öljyvahinkojätteiden käsittely Kymenlaakson alueella alusonnettomuuden jälkeen

Työn tavoitteena oli selvittää Suomenlahdella tapahtuvasta alusöljyvahingosta syntyvän öljyisen jätteen käsittelymahdollisuudet ja -kapasiteetit sekä loppusijoitusmahdollisuudet ja -kapasiteetit Kymenlaakson alueella. Lisäksi tavoitteena oli selvittää, miten öljy-vahinkojätettä voidaan esikäsitellä välivarastoinnin aikana puhdistuksen ja loppusijoituksen tehostamiseksi. Työn alussa on perehdytty öljyvahinkojätteen muodostumiseen vaikuttaviin tekijöihin: öljylaatujen ominaisuuksiin, öljyn kulkeutumiseen rannalle, ranta- ja saaristomaisemaan, öljyntorjuntaan jarantojen puhdistamiseen. Työssä on kuvattu öljyvahinkojätteen käsittelymenetelmien periaatteet ja menetelmien rajoituksia käsitellä öljyvahinkojätettä. Lisäksityö sisältää tutkimusta Suomen aluevesillä ja maailmalla tapahtuneista öljyonnettomuuksista. Onnettomuuksista on selvitetty erityisesti öljyvahinkojätteen määrä, koostumus ja käsittely. Työn loppuosassa on esitelty Kymenlaakson alueen laitosten mahdollisuuksia käsitellä öljyvahinkojätettä. Tietoa onkerätty haastattelemalla puhelimitse laitosten edustajia keväällä 2007. Alueella voidaan polttaa leijupedissä puhtaaseen polttoaineeseen sekoitettua öljyistä orgaanista ainesta ja puhdistustyössä käytettyjä varusteita arviolta 19 000 t/a, homogenoitua öljyistä orgaanista ainesta voidaan polttaa rumpu-uunissa arviolta 1200 t/a. Alueen polttokapasiteetti kasvaa, kun työn aikana rakenteilla oleva jätteenpolttolaitos valmistuu ja jätettä voidaan polttaa laitoksen arinalla. Lisäksi erityisesti öljyisiä maa-aineksia voidaan alipainekäsitellä, bitumistabiloida, kompostoida sekä pestä. Saadut tutkimustulokset ovat hyödynnettävissä erityisesti Kymenlaakson alueella. Tiedot käsittelymenetelmistä ja niiden rajoitteista ovat hyödynnettävissä valtakunnallisesti.

Ilmanpoisto kiertovoiteluöljystä

Työn kirjallisuusosassa esitellään paperi- ja kartonkikoneiden kiertovoitelujärjestelmien rakennetta ja voitelussa käytettyjen öljyjen ominaisuuksia. Lisäksi on selvitetty voiteluöljyn kunnossapidon kannalta keskeisten epäpuhtauksien kuten veden, hiukkasten ja ilmakuplien analysointia. Suurissa voitelujärjestelmissä öljyn suuri ilmapitoisuus on usein ongelma, mihin ei ole ollut selkeää ratkaisua. Työn tavoitteena oli tutkia ilmakuplien poistamista voiteluöljystä alipainekäsittelyn avulla. Alipaineen vaikusta eri öljyille ja lämpötiloilla tutkittiin laboratoriossa standarditestillä ja määritettiin sopiva alipaine tehdaskokeisiin. Testeissä havaittiin odotutetusti viskositeetin eli käytännössä lämpötilan olevan ratkaiseva tekijä ilman poistumisnopeuteen. Tehdasmittakaavan kokeissa mitattiin rakenteeltaan yksinkertaisen ja vähän energiaa kuluttavan ilmanpoistolaitteen toimintaa. Laitteisto sijoitetaan paluuöljyputkistoon ja sen ei tarvitse olla kiertovoitelukeskuksen yhteydessä. Täysimittainen laitteisto rakennettiin kartonkikoneen ja paperikoneen kiertovoitelujärjestelmiin. Laitteen avulla voidaan käsitellä koko voitelujärjestelmän öljy. Tuloksien mukaan laite toimii odotetulla tavalla ja vähentää merkittävästi ilmapitoisuutta. Järjestelmä on heikoimmillaan tilanteessa, jossa lämpötilat on pidettävä alhaisina ja ilmakuplia on runsaasti.