Maternal and fetal anaesthesia for fetal surgery.

Over the last three decades, advances in early diagnosis of fetal anomalies, imaging and surgical techniques have led to a huge expansion in fetal surgery. A small number of specialist centres perform fetal surgery, which involves high-risk anaesthesia for the mother and fetus. The anaesthetist plays an integral role within the large multispecialty and multidisciplinary team, involved in planning and delivering care for complex surgical procedures. This article reviews three fetal surgical procedures, congenital diaphragmatic hernia, myelomeningocele repair and ex-utero intrapartum treatment for airway obstruction. The underlying fetal pathology, surgical management, anaesthetic considerations and risks for both the mother and fetus are described for each. Fundamental to this is the understanding that clear communication and collaboration between all team members is vital to ensure successful outcomes of patients, the mother and the fetus.

Dietary polyunsaturated fatty acid supplementation of young post-pubertal dairy bulls alters the fatty acid composition of seminal plasma and spermatozoa but has no effect on semen volume or sperm quality.

The aim of this study was to examine the effects of dietary supplementation with rumen protected n-6 or n-3 polyunsaturated fatty acids (PUFA) on the quantity and quality of semen from young post-pubertal dairy bulls. Pubertal Holstein-Friesian (n = 43) and Jersey (n = 7) bulls with a mean ± s.e.m. age and bodyweight of 420.1 ± 5.86 days and 382 ± 8.94 kg, respectively, were blocked on breed, weight, age and semen quality (based on the outcomes of two pre-trial ejaculates) and randomly assigned to one of three treatments: (i) a non-supplemented control (CTL, n = 15), (ii) rumen-protected safflower (SO, n = 15), (iii) rumen-protected n-3 PUFA-enriched fish oil (FO, n = 20). Bulls were fed their respective diets, ad libitum for 12 weeks; individual intakes were recorded using an electronic feeding system for the initial 6 weeks of the feeding period. Semen was collected via electro-ejaculation at weeks -2, -1, 0, 7, 10, 11 and 12 relative to the beginning of the trial period (week 0). On collection, semen volume, sperm concentration and progressive linear motility (PLM) were assessed. On weeks -2, -1, 0, 10, 11, 12, semen was packaged into 0.25 mL straws and frozen using a programmable freezer. On weeks -1, 7 and 11; a sub-sample of semen was separated into sperm and seminal plasma, by centrifugation and stored at - 20 °C until analysis of lipid composition. Semen from 10 bulls per treatment were used for post-thaw analysis at weeks 10, 11 and 12 (3 straws per ejaculate). Sperm motility was analysed by computer assisted semen analysis (CASA). In addition, membrane fluidity, acrosome reaction and oxidative stress were assessed using flow cytometry. Sperm from bulls fed SO had a 1.2 fold higher total n-6 PUFA content at week 11 compared to week -1 (P < 0.01) while bulls fed FO had a 1.3 fold higher total n-3 PUFA content, in sperm by week 11 (P < 0.01). There was no effect of diet on semen volume, concentration or PLM of sperm when assessed either immediately following collection or post-thawing. Membrane fluidity and oxidative stress of sperm were also not affected by diet. The percentage of sperm with intact-acrosomes was lower in CTL bulls compared to those fed SO (P < 0.01). In conclusion, while the lipid composition of semen was altered following dietary supplementation with either n-6 or n-3 based PUFA, this did not lead to measurable improvements in the quantity or quality of semen produced by young post-pubertal dairy bulls.

Impact of sustainable feeds on omega-3 long-chain fatty acid levels in farmed Atlantic salmon, 2006-2015.

As the global population and its demand for seafood increases more of our fish will come from aquaculture. Farmed Atlantic salmon are a global commodity and, as an oily fish, contain a rich source of the health promoting long-chain omega-3 fatty acids, eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids. Replacing the traditional finite marine ingredients, fishmeal and fish oil, in farmed salmon diets with sustainable alternatives of terrestrial origin, devoid of EPA and DHA, presents a significant challenge for the aquaculture industry. By comparing the fatty acid composition of over 3,000 Scottish Atlantic salmon farmed between 2006 and 2015, we find that terrestrial fatty acids have significantly increased alongside a decrease in EPA and DHA levels. Consequently, the nutritional value of the final product is compromised requiring double portion sizes, as compared to 2006, in order to satisfy recommended EPA + DHA intake levels endorsed by health advisory organisations. Nevertheless, farmed Scottish salmon still delivers more EPA + DHA than most other fish species and all terrestrial livestock. Our findings highlight the global shortfall of EPA and DHA and the implications this has for the human consumer and examines the potential of microalgae and genetically modified crops as future sources of these important fatty acids.

Replacement of fish oil with a DHA-rich algal meal derived from Schizochytrium sp. on the fatty acid and persistent organic pollutant levels in diets and flesh of Atlantic salmon (Salmo salar, L.) post-smolts.

The replacement of fish oil (FO) with a DHA-rich Schizochytrium sp. algal meal (AM) at two inclusion levels (11% and 5.5% of diet) was tested in Atlantic salmon post-smolts compared to fish fed a FO diet of northern (NFO) or southern hemisphere (SFO) origin. Fish were preconditioned prior to the 19-week experimental feeding period to reduce long-chain polyunsaturated fatty acid (LC-PUFA) and persistent organic pollutant levels (POPs). Dietary POP levels differed significantly between treatments in the order of NFO>SFO>11 AM/5.5 AM and were subsequently reflected in the flesh. Fish fed the 11 AM diet contained similar DHA levels (g 100 g(-1) flesh) to FO-fed fish, despite percentage differences. However, the low levels of EPA in the diets and flesh of algal-fed fish compromised the overall nutritional value to the final consumer. Nevertheless, further developments in microalgae culture offer a promising alternative lipid source of LC-PUFA to FO in salmon feeds that warrants further investigation.

Lipid and fatty acid composition, and persistent organic pollutant levels in tissues of migrating Atlantic bluefin tuna (Thunnus thynnus, L.) broodstock.

Lipid class, fatty acid and POP levels were measured in migrating Atlantic bluefin tuna (ABT) tissues caught off the Barbate coast, Spain. Tissue lipids were largely characterized by triacylglycerol, reflecting large energy reserves accumulated prior to reproductive migration. Fatty acid compositions of muscle, liver and adipose exhibited similar profiles, whereas gonads showed a higher affinity for docosahexaenoic acid. Tissue POP concentrations correlated positively with percentage triacylglycerol and negatively with polar lipids. Highest POP concentrations were in adipose and lowest in gonads, reflecting lipid content. DL-PCBs contributed most to total PCDD/F + DL-PCB levels, with mono-ortho concentrations higher in tissues, whereas non-ortho PCBs contributed greater WHO-TEQs due to differences in TEFs. PBDE47 was the most prominent BDE congener in tissues, probably through biotransformation of BDE99 and other higher brominated congeners. The perceived POP risk from ABT consumption should be balanced by the well-established beneficial effects on human health of omega-3 fatty acids.

Lipid and fatty acid composition of parasitic caligid copepods belonging to the genus Lepeophtheirus.

Sea lice are copepod ectoparasites that constitute a major barrier to the sustainability and economic viability of marine finfish aquaculture operations worldwide. In particular, the salmon louse, Lepeophtheirus salmonis, poses a considerable problem for salmoniculture in the northern hemisphere. The free-swimming nauplii and infective copepodids of L. salmonis are lecithotrophic, subsisting principally on maternally-derived lipid reserves. However, the lipids and fatty acids of sea lice have been sparsely studied and therefore the present project aimed to investigate the lipid and fatty acid composition of sea lice of the genus Lepeophtheirus obtained from a variety of fish hosts. Total lipid was extracted from eggs and adult female L. salmonis obtained from both wild and farmed Atlantic salmon (Salmo salar L.) sampled at two time points, in the mid 1990s and in 2009. In addition, L. salmonis from wild sea trout (Salmo trutta L.) and L. hippoglossi from wild Atlantic halibut (Hippoglossus hippoglossus L.) were sampled and analyzed. The lipids of both females and egg strings of Lepeophtheirus were characterized by triacylglycerol (TAG) as the major neutral (storage) lipid with phosphatidylcholine and phosphatidylethanolamine as the major polar (membrane) lipids. The major fatty acids were 22:6n-3 (DHA), 18:1n-9 and 16:0, with lesser amounts of 20:5n-3, 22:5n-3 and 18:0. L. salmonis sourced from farmed salmon was characterized by higher levels of 18:2n-6 and 18:3n-3 than lice from wild salmon. Egg strings had higher levels of TAG and lower DHA compared to females, whereas L. hippoglossi had lower levels of TAG and higher DHA than L. salmonis. The results demonstrate that the fatty acid compositions of lice obtained from wild and farmed salmon differ and that changes to the lipid and fatty acid composition of feeds for farmed salmon influence the louse compositions.

Noncardiothoracic nonobstetric surgery in mild-to-moderate pulmonary hypertension.

The anaesthetic management and follow-up of well-characterised patients with pulmonary arterial hypertension presenting for noncardiothoracic nonobstetric surgery has rarely been described. The details of consecutive patients and perioperative complications during the period January 2000 to December 2007 were reviewed. Repeat procedures in duplicate patients were excluded. Longer term outcomes included New York Heart Association (NYHA) functional class, 6-min walking distance and invasive haemodynamics. A total of 28 patients were identified as having undergone major (57%) or minor surgery under general (50%) and regional anaesthesia. At the time of surgery, 75% of patients were in NYHA functional class I-II. Perioperative deaths occurred in 7%. Perioperative complications, all related to pulmonary hypertension, occurred in 29% of all patients and in 17% of those with no deaths during scheduled procedures. Most (n = 11, 92%) of the complications occurred in the first 48 h following surgery. In emergencies (n = 4), perioperative complication and death rates were higher (100 and 50%, respectively; p<0.005). Risk factors for complications were greater for emergency surgery (p<0.001), major surgery (p = 0.008) and a long operative time (193 versus 112 min; p = 0.003). No significant clinical or haemodynamic deterioration was seen in survivors at 3-6 or 12 months of post-operative follow-up. Despite optimal management in this mostly nonsevere pulmonary hypertension population, perioperative complications were common, although survivors remained stable. Emergency procedures, major surgery and long operations were associated with increased risk.

The effect of peripheral nerve injury on disease progression in the SOD1(G93A) mouse model of amyotrophic lateral sclerosis.

Around 20% of familial cases of amyotrophic lateral sclerosis have been shown to carry mutations in Cu/Zn superoxide dismutase 1 (Cu/Zn SOD1). Transgenic mice over-expressing human mutant SOD1 genes have been developed and in this study we examined the effect of nerve injury on disease progression in these mice. Firstly, disease progression in uninjured mice was characterised using physiological methods. Muscle force, contractile characteristics and motor unit survival was established at 90 days, an early symptomatic stage and also at the end-stage of the disease, at 130 days. In addition, muscle histochemistry was examined and the extent of motoneuron survival established morphologically. By 90 days of age, there is a significant reduction in muscle force, and nearly 40% of motoneurons within the sciatic motor pool have already died. By 130 days, the muscles are significantly weaker, and there is a dramatic change in the phenotype of extensor digitorum longus (EDL), which changes from a fast fatigable muscle, to a fatigue resistant muscle with a high oxidative capacity. By this stage of the disease, only 40% of motor units in EDL survive, with only 29% of motoneurons surviving within the sciatic motor pool. Following injury to the sciatic nerve in SOD1(G93A) mice, there is an acceleration in disease progression so that 90 day old mice show deficits that are only seen at the end stage in uninjured SOD1(G93A) mice. It is therefore possible that mutant SOD1 toxicity increases the vulnerability of motoneurons and muscles to stressful stimuli such as nerve injury.

Essential fatty acids and phospholipase A2 in autistic spectrum disorders.

A health questionnaire based on parental observations of clinical signs of fatty acid deficiency (FAD) showed that patients with autism and Asperger's syndrome (ASP) had significantly higher FAD scores (6.34+/-4.37 and 7.64+/-6.20, respectively) compared to controls (1.78+/-1.68). Patients with regressive autism had significantly higher percentages of 18:0,18:2n-6 and total saturates in their RBC membranes compared to controls, while 24:0, 22:5n-6, 24:1 and the 20:4n-6/20:5n-3 ratio were significantly higher in both regressive autism and ASP groups compared to controls. By comparison, the 18:1n-9 and 20:4n-6 values were significantly lower in patients with regressive autism compared to controls while 22:5n-3, total n-3 and total dimethyl acetals were significantly lower in both regressive autism and ASP groups compared to controls. Storage of RBC at -20 degrees C for 6 weeks resulted in significant reductions in highly unsaturated fatty acid levels in polar lipids of patients with regressive autism, compared to patients with classical autism or ASP, or controls. Patients diagnosed with both autism and ASP showed significantly increased levels of EPA ( approximately 200%) and DHA ( approximately 40%), and significantly reduced levels of ARA ( approximately 20%), 20:3n-6 and ARA/EPA ratio in their RBC polar lipids, when supplemented with EPA-rich fish oils, compared to controls and non-supplemented patients with autism. Patients with both regressive autism and classical autism/Asperger's syndrome had significantly higher concentrations of RBC type IV phospholipase A2 compared to controls. However, patients with autism/ASP, who had taken EPA supplements, had significantly reduced PLA2 concentrations compared to unsupplemented patients with classical autism or ASP.

Over-expression of parvalbumin in transgenic mice rescues motoneurons from injury-induced cell death.

Following nerve injury in neonatal rats, a large proportion of motoneurons die, possibly as a consequence of an increase in vulnerability to the excitotoxic effects of glutamate. Calcium-dependent glutamate excitotoxicity is thought to play a significant role not only in injury-induced motoneuron death, but also in motoneuron degeneration in diseases such as amyotrophic lateral sclerosis (ALS). Motoneurons are particularly vulnerable to calcium influx following glutamate receptor activation, as they lack a number of calcium binding proteins, such as calbindin-D(28k) and parvalbumin. Therefore, it is possible that increasing the ability of motoneurons to buffer intracellular calcium may protect them from cell death and prevent the decline in motor function that usually occurs as a consequence of motoneuron loss. In this study we have tested this possibility by examining the effect of neonatal axotomy on motoneuron survival and muscle force production in normal and transgenic mice that over-express parvalbumin in their motoneurons.The sciatic nerve was crushed in one hindlimb of new-born transgenic and wildtype mice. The effect on motoneuron survival was assessed 8 weeks later by retrograde labelling of motoneurons innervating the tibialis anterior muscle. Following nerve injury in wildtype mice, only 20.2% (+/-2.2, S.E.M.; n=4) of injured motoneurons survive long term compared with 47.2% (+/-4.4, S.E.M.; n=4) in parvalbumin over-expressing mice. Surprisingly, this dramatic increase in motoneuron survival was not reflected in a significant improvement in muscle function, since 8 weeks after injury there was no improvement in either maximal twitch and tetanic force, or muscle weights.Thus, inducing spinal motoneurons to express parvalbumin protects a large proportion of motoneurons from injury-induced cell death, but this is not sufficient to restore muscle function.

Pyloric ceca are significant sites of newly synthesized 22:6n-3 in rainbow trout (Oncorhynchus mykiss).

In this pulse-chase study, rainbow trout fed a diet containing deuterated (D5) (17,17,18,18,18)-18:3n-3 ethyl ester accumulated D5-22:6n-3 in pyloric ceca to a greater extent than in liver 2 d post-dose. The ratio of newly synthesized D5-22:6n-3 in ceca to that in liver 2 d after feeding D5-18:3n-3 was 4.7 +/- 1.2 when expressed as per mg tissue and 5.2 +/- 2.4 when expressed as per mg protein. The amount of D5-22:6n-3 in ceca then declined whereas that in liver and blood increased, with the ratio of ceca to liver falling to 1.7 and 1.4, respectively, by day 5 and approaching unity by day 9. A crude cecal mucosa fraction contained 123 +/- 50 ng D5-22:6n-3/mg protein/mg D5-18:3n-3 eaten 2 d after feeding the tracer, compared with 35 +/- 21 ng D5-22:6n-3/mg protein/mg D5-18:3n-3 eaten in liver. Three days later the amount in cecal mucosa had fallen by one-third and that in liver had increased threefold. Most of the D5-18:3n-3 was catabolized very rapidly. The ratio of D5-18:3n-3 to 21:4n-6 (a relatively inert FA marker) in the diet was 4.0, but this fell to 0.30 in ceca and ca. 0.8 in liver, blood, and whole carcass one day after feeding. These results indicate that ceca are active in the synthesis of 22:6n-3 and the oxidation of 18:3n-3.

A vertebrate fatty acid desaturase with Delta 5 and Delta 6 activities.

Delta5 and Delta6 fatty acid desaturases are critical enzymes in the pathways for the biosynthesis of the polyunsaturated fatty acids arachidonic, eicosapentaenoic, and docosahexaenoic acids. They are encoded by distinct genes in mammals and Caenorhabditis elegans. This paper describes a cDNA isolated from zebrafish (Danio rerio) with high similarity to mammalian Delta6 desaturase genes. The 1,590-bp sequence specifies a protein that, in common with other fatty acid desaturases, contains an N-terminal cytochrome b(5) domain and three histidine boxes, believed to be involved in catalysis. When the zebrafish cDNA was expressed in Saccharomyces cerevisiae it conferred the ability to convert linoleic acid (18:2n-6) and alpha-linolenic acid (18:3n-3) to their corresponding Delta6 desaturated products, 18:3n-6 and 18:4n-3. However, in addition it conferred on the yeast the ability to convert di-homo-gamma-linoleic acid (20:3n-6) and eicosatetraenoic acid (20:4n-3) to arachidonic acid (20:4n-6) and eicosapentaenoic acid (20:5n-3), respectively, indicating that the zebrafish gene encodes an enzyme having both Delta5 and Delta6 desaturase activity. The zebrafish Delta5/Delta6 desaturase may represent a component of a prototypic vertebrate polyunsaturated fatty acids biosynthesis pathway.

Hepatocyte fatty acid desaturation and polyunsaturated fatty acid composition of liver in salmonids: effects of dietary vegetable oil.

The desaturation and elongation of [1-(14)C]18:3n-3 was investigated in hepatocytes from different populations and three different species of salmonids indigenous to Scotland, brown trout, Atlantic salmon and Arctic charr. Two groups of fish were sampled, before and after they were fed two experimental diets, a control diet containing fish oil and a diet containing vegetable oil (a 1:1 blend of linseed and rapeseed oils) for 12 weeks. At each sampling time, fatty acyl desaturation and elongation activity was determined in isolated hepatocytes, and samples of liver were also collected for lipid compositional analysis. At the initiation of the dietary trial, the liver polar-lipid fatty acid compositions of salmon and brown trout were very similar to each other, and the two charr populations were similar to each other, having lower total n-3 polyunsaturated fatty acids (PUFA) and 22:6n-3, but higher 20:5n-3 than the other salmonids. Initially, hepatocyte desaturation activity varied, with the highest activity in brown trout, followed by salmon and then charr. Production of 20:5n-3 was particularly high in brown trout. Desaturation of [1-(14)C]18:3n-3 was significantly greater in all fish fed the diet containing vegetable oil compared to fish fed the diet containing fish oil. The increase in activity was less in brown trout compared to the other groups of fish. Feeding the vegetable oil diet increased the levels of 18:2n-6, 20:3n-6, total n-6 PUFA, 18:3n-3, 18:4n-3, 20:3n-3 and 20:4n-3, and decreased 22:6n-3 and the n-3/n-6 ratio in salmon and brown trout. By contrast, in charr fed the vegetable oil diet, there was no increase in 18:3n-3, 18:4n-3, 20:3n-3 or 20:4n-3 in liver polar lipids and the level of 22:6n-3 was not decreased. In addition, there was only a modest increase in the levels of 18:2n-6 and total n-6 PUFA, and so the n-3/n-6 ratio was only slightly decreased. The percentage of 20:4n-6, which was not increased in salmon and brown trout fed vegetable oil, was increased in charr fed the vegetable oil diet. Overall, the results indicated that there were significant differences in liver PUFA metabolism between Arctic charr and the other salmonids, which could have important consequences, both physiologically and in their ability to be successfully cultured on diets containing vegetable oils.

Effects of essential fatty acid deficiency and supplementation with docosahexaenoic acid (DHA; 22:6n-3) on cellular fatty acid compositions and fatty acyl desaturation in a cell culture model.

The desaturation of [1-(14)C] 18:3n-3 to docosahexaenoic acid (DHA; 22:6n-3) is enhanced in an essential fatty acid deficient cell line (EPC-EFAD) in comparison with the parent cell line (EPC) from carp. In the present study, the effects of DHA on lipid and fatty acid compositions, and the metabolism of [1-(14)C]18:3n-3 were investigated in EPC-EFAD cells in comparison with EPC cells. DHA supplementation had only relatively minor effects on lipid content and lipid class compositions in both EPC and EPC-EFAD cells, but significantly increased the amount of DHA, 22:5n-3, eicosapentaenoic acid (EPA; 20:5n-3), total n-3 polyunsaturated fatty acids (PUFA), total PUFA and saturated fatty acids in total lipid and total polar lipid in both cell lines. Retroconversion of supplemental DHA to EPA was significantly greater in EPC cells. Monounsaturated fatty acids, n-9 and n-6PUFA were all decreased in total lipid and total polar lipid in both cell lines by DHA supplementation. The incorporation of [1-(14)C]18:3n-3 was greater into EPC-EFAD compared to EPC cells but DHA had no effect on the incorporation of [1-(14)C]18:3n-3 in either cell line. In contrast, the conversion of [1-(14)C]18:3n-3 to tetraenes, pentaenes and total desaturation products was similar in the two cell lines and was significantly reduced by DHA supplementation in both cell lines. However, the production of DHA from [1-(14)C]18:3n-3 was significantly greater in EPC-EFAD cells compared to EPC cells and, whereas DHA supplementation had no effect on the production of DHA from [1-(14)C]18:3n-3 in EPC cells, DHA supplementation significantly reduced the production of DHA from [1-(14)C] 18:3n-3 in EPC-EFAD cells. Greater production of DHA in EPC-EFAD cells could be a direct result of significantly lower levels of end-product DHA in these cells' lipids compared to EPC cells. Consistent with this, the suppression of DHA production upon DHA supplementation was associated with increased cellular and membrane DHA concentrations in EPC-EFAD cells. However, an increase in cellular DHA content to similar levels failed to suppress DHA production in DHA-supplemented EPC cells. A possible explanation is that greatly increased levels of EPA, derived from retroconversion of the added DHA, acts to offset the suppression of the pathway by DHA by stimulating conversion of EPA to DHA in DHA-supplemented EPC cells.

Biosynthesis of eicosapentaenoic acid in the sea urchin Psammechinus miliaris.

The sea urchin Psammechinus miliaris (Gmelin) (Echinodermata: Echinoidea) was shown by using a deuterated tracer (D5-18:3n-3) and quantitation by negative chemical ionization gas chromatography-mass spectrometry to convert 18:3n-3 to 20:5n-3. The rate of conversion was very slow, corresponding to 0.09 microg/g tissue/mg 18:3n-3 eaten over 14 d. Deuterated arachidonic acid (D8-20:4n-6) was also included in the diet to give a measure of the relative amounts of diet eaten by the different animals. The recovery of this fatty acid in tissue lipids was 33.7% compared with only 0.95% recovery of D5-18:3n-3 and its anabolites, indicating that the majority of the D5-tracer was catabolized. Considerable elongation of D5-18:3n-3 into 20:3n-3 and a trace of 22:3n-3 was found, and these were accompanied by minor amounts of the intermediates 18:4n-3 and 20:4n-3. No deuterated 22:6n-3 was found.

Biosynthesis and tissue deposition of docosahexaenoic acid (22:6n-3) in rainbow trout (Oncorhynchus mykiss).

Rainbow trout (Oncorhynchus mykiss) weighing ca. 5 g and previously acclimated for 8 wk on a diet comprising vegetable oil (11%), fish meal (5%), and casein (48%) as the major constituents were fed a pulse of diet containing deuterated (D5) (17,17,18,18,18)-18:3n-3 ethyl ester. The synthesis and tissue distribution of D5-22:6n-3 was determined 3, 7, 14, 24, and 35 d after the pulse. The whole-body accumulation of D5-22:6n-3 was linear over the first 7 d, corresponding to a rate of 0.54 +/- 0.12 microg D5-22:6n-3/g fish/mg D5-18:3n-3 eaten/d. Maximal accretion of D5-22:6n-3 was 4.3 +/- 1.2 microg/g fish/mg of D5-18:3n-3 eaten after 14 d. The amount of D5-22:6n-3 peaked in liver at day 7, in brain and eyes at day 24, and plateaued after day 14 in visceral and eye socket adipose tissue and in the whole fish. The majority of D5-22:6n-3 was found in the carcass (remaining tissues minus the above tissues analyzed separately) at all times. On a per milligram lipid basis, liver and eyes had the highest concentration of D5-22:6n-3. The experimental diet also contained 21:4n-6 ethyl ester as a marker to estimate the amount of food eaten by individual fish. From such estimates it was calculated that the great majority of the D5-tracer was catabolized, with the combined recovery of D5-18:3n-3 plus D5-22:6n-3 being 2.6%. The recovery of 21:4n-6 was 57.6%. The concentration of 22:6n-3 in the fish decreased during the 13-wk period, and the amount of 22:6n-3 synthesized from 18:3n-3 was only about 5% of that obtained directly from the fish meal in the diet.

Red blood cell fatty acid compositions in a patient with autistic spectrum disorder: a characteristic abnormality in neurodevelopmental disorders?

The fatty acid compositions of red blood cell (RBC) phospholipids from a patient with autistic spectrum disorder (ASD) had reduced percentages of highly unsaturated fatty acids (HUFA) compared to control samples. The percentage of HUFA in the RBC from the autistic patient was dramatically reduced (up to 70%) when the sample was stored for 6 weeks at -20 degrees C. However, only minor HUFA reductions were recorded in control samples stored similarly, or when the autistic sample was stored at -80 degrees C. A similar instability in RBC HUFA compositions upon storage at -20 degrees C has been recorded in schizophrenic patients. In a number of other neurodevelopmental conditions, including attention deficit hyperactivity disorder (ADHD) and dyslexia, reduced concentrations of RBC HUFA have been recorded. The extent and nature of these aberrations require further assessment to determine a possible common biochemical origin of neurodevelopmental disorders in general. To facilitate this, a large scale assessment of RBC fatty acid compositions in patients with ASD, and related disorders, should be performed as a matter of urgency. Supplementing cells in culture with the tryptophan metabolite indole acrylic acid (IAA) affected the levels of cellular HUFA and prostaglandin production. Indole acroyl glycine (IAG), a metabolite of IAA excreted in urine, is found in high concentrations in patients with neurodevelopmental disorders including ASD, ADHD, dyslexia, Asperger's syndrome and obsessive compulsive disorder.

Integrins at the neuromuscular junction are important for motoneuron survival.

During development motoneurons depend on target contact for their survival. Following injury to the sciatic nerve in neonatal rats, a large proportion of motoneurons die. However, the same injury inflicted at 5 days of age results in no loss of motoneurons. This critical period of postnatal development coincides with the time during which there is a significant increase in the release of transmitter from the nerve terminals at the neuromuscular junction. We have proposed that the role of the target muscle cell during this period is to induce this up-regulation of transmitter release from motor nerve terminals. It has been shown that stretch-induced increase in transmitter release from frog motor nerve terminals is accomplished via an integrin-dependent mechanism. In this study we examined the role of integrins at the rat neuromuscular junction in motoneuron survival. We found that blocking integrin binding at the developing neuromuscular junction delayed the increase in choline acetyltransferase activity that normally takes place during the early postnatal period, and resulted in motoneuron death. Furthermore, the maturation of those motoneurons that survived was delayed so they remained susceptible to subsequent nerve injury. These results support the possibility that integrins, by their involvement in modulating transmitter release, can influence motoneuron survival.

Low C18 to C20 fatty acid elongase activity and limited conversion of stearidonic acid, 18:4(n-3), to eicosapentaenoic acid, 20:5(n-3), in a cell line from the turbot, Scophthalmus maximus.

The TF cell line, derived from a top predatory, carnivorous marine teleost, the turbot (Scophthalmus maximus), is known to have a limited conversion of C18 to C20 polyunsaturated fatty acids (PUFA). To illuminate the underlying processes, we studied the conversions of stearidonic acid, 18:4(n-3), and its elongation product, 20:4(n-3), in TF cells and also in a cell line, AS, derived from Atlantic salmon (Salmo salar), by adding unlabelled (25 microM), U-14C (1 microM) or deuterated (d5; 25 microM) fatty acids. Stearidonic acid, 18:4(n-3), was metabolised to 20:5(n-3) in both cells lines, but more so in AS than in TF cells. Delta5 desaturation was more active in TF cells than in AS cells, whereas C18 to C20 elongation was much reduced in TF as compared to AS cells. Only small amounts of docosahexaenoic acid (22:6(n-3)) were produced by both cell lines, although there was significant production of 22:5(n-3) in both cultures, especially when 20:4(n-3) was supplemented. We conclude that limited elongation of C18 to C20 fatty acids rather than limited fatty acyl Delta5 desaturation accounts for the limited rate of conversion of 18:3(n-3) to 20:5(n-3) in the turbot cell line, as compared to the Atlantic salmon cell line. The results can account for the known differences in conversions of C18 to C20 PUFA by the turbot and the Atlantic salmon in vivo.

Molecular speciation of fish sperm phospholipids: large amounts of dipolyunsaturated phosphatidylserine.

The molecular species compositions of the main diacyl phosphoglyceride classes and ether-linked subclasses from sperm of three species of fish, sea bass Dicentrarchus labrax, Atlantic salmon Salmo salar and Chinook salmon Onchorhynchus tsawytscha, were determined. The phospholipids from sperm were highly unsaturated, dipolyunsaturated fatty acid (diPUFA) molecular species comprised 64.6 to 71.8% of phosphatidylserine (PS), 10.1 to 17.4% of phosphatidylethanolamine (PE), and 3.3 to 10.1% of phosphatidylcholine (PC). In sea bass sperm, di22:6n-3 phospholipid was the predominant diPUFA molecular species, but in both salmon species 22:5n-3/22:6n-3 was also a major constituent of PS. Phospholipids containing 22:6n-3 dominated in sea bass sperm with 16:0/22:6n-3 as a major component of PC and PE, and 18:0/22:6n-3 of PE and PS in addition to di22:6n-3 in the latter two classes. In contrast, both salmon species contained much more 20:5n-3 and less 22:6n-3 so that saturated/20:5n-3 and monounsaturated/20:5n-3 molecular species were more abundant than the corresponding molecules containing 22:6n-3. Ether-linked lipids comprised 11.3-36.3% of choline and ethanolamine phosphoglycerides in each fish species. Molecular species containing 22:6n-3 were the major components of 1-O-alkyl-2-acyl-glycerophosphocholine, especially 16:0a/22:6n-3 in sea bass and 18:1a/22:6n-3 in the two salmon species, while in 1-O-alk-1'-enyl-2-acyl-glycerophosphoethanolamine, 16:0a/22:6n-3 was the major component in both salmon and 18:0a/22:6n-3 in sea bass with 18:1a/22:6n-3 abundant in all three species. In Atlantic salmon 1-O-alkyl-2-acyl-glycerophosphoethanolamine comprised 24.6% of ethanolamine glycerophospholipids which were predominantly 16:0a/22:6n-3 and 18:1a/22:6n-3. Phosphatidylinositol from sperm was dominated by stearoyl/C20 PUFA molecular species, in sea bass overwhelmingly 18:0/20:4n-6, while in both salmon species 18:0/20:4n-6 and 18:0/20:5n-3 were equally abundant.