Molecular epidemiology of methicillin-resistant Staphylococcus aureus in Israel: dissemination of global clones and unique features.

From 2006 to 2009, 315 clinical methicillin-resistant Staphylococcus aureus (MRSA) isolates were collected from 5 hospitals across Israel. Most isolates (64%) were related to the global clones spa types t001-SCCmec-I (SCCmec-I stands for staphylococcal cassette chromosome mec type I) (n = 99; 31%), t002-SCCmec-II (n = 82; 26%), and t008-SCCmec-IV (n = 21; 7%), five of which were identified as MRSA strain USA-300. Seventeen strains unique to Israel were identified. SCCmec types IV and V were common among hospital-acquired isolates.

Mucormycosis in a liver allograft: salvage re-transplantation and targeted immunosuppressive management.

Zygomycetes infection is associated with a high mortality in transplant populations. We describe a child with liver allograft Rhizopus oryzae infection who was salvaged by liver re-transplantation. A 10-year-old child presented with anastomotic bile leak that was repaired. A combined antibiotics and voriconazole regimen was introduced for Escherichia coli and Candida krusei growth in the peritoneal fluid. Despite broad antibiotic and antifungal coverage, the patient continued to have an ongoing infection. A follow-up computed tomography scan 8 weeks later showed 2 liver abscesses infiltrating the stomach and the diaphragm, with splenic infarcts and pericardial effusion. Aspirated samples from the liver abscess and the pericardial fluid revealed R. oryzae. Immunosuppression was discontinued and an antifungal regimen combining amphotericin B, posaconazole, and caspofungin was introduced. After 3 weeks of treatment with control of the systemic signs of infection, a positron emission tomography showed the fluorescence stain limited to the liver. With infection confined to the liver, the child underwent liver re-transplantation, splenectomy, and partial gastrectomy. Immunosuppression was reintroduced with recovery of the immune response observed by the CD4 cells adenosine triphophate release (Cylex(™) ImmuKnow(®) assay) and posaconazole was continued for another year. At 3-year follow-up, the child maintained normal graft function. We conclude that discontinuation of immunosuppression combined with a modern antifungal regimen may allow salvage re-transplantation in patients with liver mucormycosis.

Chronic granulomatous disease of childhood: an unusual cause of recurrent uncommon infections in a 61-year-old man.

Chronic granulomatous disease (CGD) is a rare congenital immunodeficiency that affects 1 : 250,000 of the population, which is characterized by recurrent bacterial and fungal infections and by granuloma formation. We investigated a 61-year-old man presented with a 20-year history of a relapsing skin rash appearing as mildly pruritic and erythematous plaques affecting various body regions. Cutaneous biopsies were taken and sent for histology and tissue culture. Leucocyte function was assessed by determining the generation of reactive oxygen species. Bactericidal activity was assessed in the presence of autologous and homologous sera. Western blotting was performed for protein analysis of the reduced nicotinamide adenine dinucleotide phosphate oxidase system, and mutation screening was carried out using PCR amplification and sequence analysis. Examination of biopsies obtained from lesional skin indicated a suppurative granulomatous process. Tissue cultures grew Aspergillus nidulans and Aspergillus fumigatus (confirmed by PCR). A. nidulans has often been associated with CGD, and the leucocyte function tests supported this diagnosis. Direct DNA sequencing led to the identification of a hemizygous missense novel mutation in CYBB (c.907C>T), which predicts a p.His303Tyr amino-acid substitution in gp91-phox, thus confirming the diagnosis of CGD. In conclusion, we report a case of a rare inherited immunodeficiency, CGD, in a 61-year-old man, and describe the novel hemizygous missense mutation underlying the condition. Mild forms of usually fatal immunodeficiencies should be considered when assessing the occurrence of unusual infectious diseases in apparently healthy people.

Radiological findings of early invasive pulmonary aspergillosis in immune-compromised patients.

Data on the radiological features of invasive pulmonary aspergillosis (IPA) in early stages is scanty. Detection of Aspergillus (ASP) species in broncho-alveolar (BAL) fluid by polymerase chain reaction (PCR) enables early diagnosis of IPA. This study describes the radiological features of early stages of IPA. Chest computerized tomography (CT) films of 22 consecutive immune-compromised patients with IPA diagnosed with the aid of ASP PCR testing from BAL fluid were characterized and compared to that of 18 similar patients diagnosed with traditional bacteriological methods and to data from the literature. It was found that patients diagnosed with the aid of ASP PCR testing tended to have focal disease as manifested by more 11-30 mm nodules with halo (68% vs. 33%, p = 0.04), more focal ground glass (single area 32% vs. 6%, p = 0.05, patchy 32% vs. 0%, p = 0.01) and less diffuse ground glass (0% vs. 22%, p = 0.03), less cavitations (5% vs. 28%, p = 0.05) and less consolidations (segmental 14% vs. 50%, p = 0.02 and diffuse 14% vs. 67%, p = 0.001). It was concluded that the radiological appearance of early IPA diagnosed with the aid of PCR testing included mainly discrete small nodules with halo and focal ground glass, representing the early stage of the disease.

Clinical impact of a PCR assay for rapid identification of Klebsiella pneumoniae in blood cultures.

The clinical impact of a rapid PCR identification assay for Klebsiella pneumoniae in positive blood cultures was prospectively evaluated. Multivariate analysis identified the rapid PCR assay as the only significant factor in decreasing the time lapse preceding the initiation of appropriate antimicrobial therapy (hazards ratio, 3.03; confidence interval, 1.62 to 5.68; P, 0.001).

High affinity esterification of eicosanoid precursor fatty acids by platelets.

We have examined the relative rates of uptake of several fatty acids into washed, human platelets by measuring incorporation into cellular phospholipids. In the presence of 15 microM fatty acid-free albumin and with radioactive fatty acid concentrations of 5-500 nM, esterification into phospholipid was linear with time and platelet concentration and saturable with respect to fatty acid concentration. Two distinct classes of uptake rate were observed. Arachidonate and 5,8,11,14,17-eicosapentaenoate exhibited high affinity, relatively rapid incorporation into platelet phospholipids at pH 6.5: apparent Michaelis constant (Km) = 30 nM, apparent maximum velocity (Vmax) = 28 pmol/min per 10(9) platelets. Two other eicosanoid precursors, 5,8,11-eicosatrienoate and 8,11,14-eicosatrienoate, exhibited the same Vmax, but Km of 85 and 60 nM, respectively. Under the same conditions, stearate, oleate, and linoleate were incorporated into phospholipids much less efficiently (Vmax approximately 8 pmol/10(9) cells per min, apparent Km greater than or equal to 170 nM). Qualitatively similar results were found at pH 7.4. Uptake of radiolabeled, rapid-uptake fatty acids was not diminished by the presence of excess, unlabeled, slow-uptake fatty acids. Thus, the specificity of this esterification system resembles that of the arachidonate-specific, long-chain acyl-CoA synthetase present in platelets. It may represent the expression in vivo of the synthetase, although the apparent affinity of the synthetase for fatty acid is much less. This esterification system probably represents the physiologic mechanism for platelet arachidonate uptake, whereby arachidonate is collected from plasma, despite the fact that its concentration is considerably lower than that of other plasma fatty acids.

2,4-Dienoyl-coenzyme A reductase deficiency: a possible new disorder of fatty acid oxidation.

Several inherited disorders of fatty acid beta-oxidation have been described that relate mainly to saturated precursors. This study is the first report of an enzyme defect related only to unsaturated fatty acid oxidation and provides the first in vivo evidence that fat oxidation in humans proceeds by the reductase-dependent pathway. The patient was a black female, presenting in the neonatal period with persistent hypotonia. Biochemical studies revealed hyperlysinemia, hypocarnitinemia, normal organic acid profile, and an unusual acylcarnitine species in both urine and blood. The new metabolite was positively identified by mass spectrometry as 2-trans,4-cis-decadienoylcarnitine, derived from incomplete oxidation of linoleic acid. In spite of dietary therapy, the patient died of respiratory acidosis at four months of age. Samples of liver and muscle from the autopsy were assayed for 2,4-dienoyl-coenzyme A reductase activity. Using the substrate 2-trans,4-cis-decadienoylcoenzyme A, the reductase activity was 40% of the control value in liver and only 17% of that found in normal muscle. It is suggested that unsaturated substrates should be used for in vitro testing to cover the full range of potential beta-oxidation defects and that acylcarnitine species identification be used for in vivo detection of this disorder.

A prospective randomized trial of itraconazole vs fluconazole for the prevention of fungal infections in patients with acute leukemia and hematopoietic stem cell transplant recipients.

Fluconazole antifungal prophylaxis is standard care in allogeneic hematopoietic stem cell transplant (HSCT) recipients, but this drug lacks anti-Aspergillus activity, the primary cause of invasive fungal infection (IFI) in many transplantation centers. We performed a randomized trial to compare itraconazole vs fluconazole, for prevention of IFIs in patients with acute leukemia (AL) and HSCT recipients. One hundred and ninety-five patients were randomly assigned to either fluconazole or itraconazole antifungal prophylaxis, after stratification into high-risk and low-risk groups. Antifungal prophylaxis was started at the beginning of chemotherapy and continued until resolution of neutropenia, or until amphotericin B treatment was started. IFI occurred in 11 (11%) of itraconazole, and in 12 (12%) fluconazole recipients. Invasive candidiasis (IC) developed in two (2%) itraconazole and one (1%) fluconazole recipients, while invasive aspergillosis (IA) developed in nine (9%) itraconazole and 11(11%) fluconazole recipients. There was no difference in the incidence of total IFI, IC and IA between the two study arms. However, there was a nonsignificant trend towards reduced mortality among patients who developed IA while receiving itraconazole prophylaxis (3/9=33% vs 8/11=73%, P=0.095).

The metabolism of (n-3) and (n-6) fatty acids and their oxygenation by platelet cyclooxygenase and lipoxygenase.

Arachidonic acid is the principal unsaturated acid in most membrane lipids. Membrane lipids also contain a variety of other (n-6) and (n-3) fatty acids. The amounts of (n-6) and (n-3) fatty acids in membrane lipids can be modified by dietary fat change. Our studies show that long chain (n-6) and (n-3) acids are metabolized by platelet lipoxygenase and cyclooxygenase. When cells are exposed to various agonists, a variety of unsaturated fatty acids may be released. Our studies show that they have the potential of modifying physiological function both by mediating arachidonic acid metabolism and as direct precursors for oxygenated metabolites which themselves may interact with specific receptors to regulate biological processes.

On the mechanistic reasons for the dual positional specificity of the reticulocyte lipoxygenase.

A set of octadecadienoic acid isomers and selected eicosatrienoic acids were tested as substrates for the lipoxygenases from soybeans and reticulocytes. Among the dienoic fatty acids, 8Z,11Z-octadecadienoic acid containing a n - 9 doubly allylic methylene group turned out to be the best substrate for the reticulocyte enzyme. This substrate was converted to its corresponding n - 7 hydroperoxy derivative. The soybean lipoxygenase, in contrast, prefers the 9Z,12Z-octadecadienoic acid (linoleic acid) which is oxygenated to its n - 6 hydroperoxy derivative. In both cases a strong preference for the LS-isomer has been observed. Analysis of the oxygenation products formed from various eicosatrienoic acids indicated that 8Z,11Z,14Z-eicosatrienoic acid was converted by the reticulocyte enzyme to its 12S- and 15S-hydroperoxy derivative in a ratio of about 1:7 (dual positional specificity), whereas the 7Z,10Z,13Z-isomer was oxygenated predominantly (greater than 97%) to its 14S-hydroperoxy derivative (singular positional specificity). 9Z,12Z,15Z-eicosatrienoic acid was oxygenated with a dual positional specificity to the corresponding 13- and 16-hydroperoxy compounds in a ratio of about 7:1. The soybean lipoxygenase converts the 8Z,11Z,14Z-isomer with a singular positional specificity to the corresponding 15S-hydroperoxy derivatives. The 9Z,12Z,15Z-eicosatrienoic acid, however, was oxygenated with a dual positional specificity to its 13S-hydroperoxy and 16S-hydroperoxy derivative in a ratio of about 1:4.

A comparison of the specific activities of linoleate and arachidonate in liver, heart and kidney phospholipids after feeding rats ethyl linoleate-9,10,12,13-d4.

In order to determine how dietary linoleate is metabolized, rats were maintained on a chemically defined diet containing 1.6% ethyl linoleate. After 5 weeks the linoleate was replaced by an equal amount of ethyl 9,10,12,13-d4-linoleate. The animals were killed 3 days later and the molar percentage of d4-linoleate and d4-arachidonate were quantified in liver, heart and kidney phospholipids. In liver, 54 and 22.8 mol% respectively of the esterified linoleate and arachidonate was deuteriated. The lower specific activity of arachidonate versus linoleate suggests that desaturation of linoleate, by a 6-desaturase, is not only rate limiting for synthesis of arachidonate but that the amount of newly synthesized arachidonate is insufficient by itself to maintain steady state levels of esterified arachidonate. The molar fraction of deuteriated linoleate in heart and kidney phospholipids was respectively 35 and 37.4%. These values are lower than for liver phospholipids but it appears there is adequate dietary linoleate available in these tissues for the synthesis of arachidonate. However, of the esterified arachidonate in heart and kidney phospholipids only 4.2 and 8.6 mol% respectively was deuteriated. Our results suggest that arachidonate is made in liver and transported to heart and kidney.

Studies to determine if rat liver contains multiple chain elongating enzymes.

According to the revised pathways of polyunsaturated fatty acid biosynthesis three, rather than two acids, must be chain elongated for converting linoleate and linolenate, respectively, to 22:5(n-6) and 22:6(n-3) (Sprecher et al. (1995) J. Lipid Res. 36, 2471-2477). The present study was undertaken to determine whether microsomes contained chain-length specific chain-elongating enzymes and, secondly, whether reaction rates for any of these reactions might be rate limiting in the synthesis of 24:5(n-6) and 24:6(n-3), which are the immediate precursors of 22:5(n-6) and 22:6(n-3). Rates of total chain elongation products produced from both 18:4(n-3) and 20:5(n-3) were about 3 nmol/min/mg of microsomal protein while only about 0.5 nmol/min/mg of 24:5(n-3) plus 24:6(n-3) was synthesized from 22:5(n-3). The rate of 24:5(n-3) synthesis was similar to that for the desaturation of 24:5(n-3), at position 6, to yield 24:6(n-3) (Geiger et al. (1993) Biochim. Biophys. Acta 1170, 137-142). The results suggest that the last chain elongation step in unsaturated fatty acid biosynthesis may be equally regulatory in governing the synthesis of fatty acids as is desaturation at position 6. When an enzyme saturating level of [1-(14)C]18:4(n-3) was incubated with increasing amounts of 18:3(n-6) there was a decrease in the production [1-(14)C]20:4(n-3). In a similar way it was observed that 18:4(n-3) inhibited the chain elongation of [1-(14)C]18:3(n-6). Identical cross-over inhibitory studies, using 20:4(n-6) and 20:5(n-3), as well as 22:4(n-6) and 22:5(n-3) also suggested that microsomes contain chain length specific chain-elongating enzymes. This conclusion was further supported by the finding that neither 20:5(n-3) or 22:5(n-3) inhibited the chain elongation of [1-(14)C]18:4(n-3). However, 18:4(n-3), and to a lesser degree, 22:5(n-3) did inhibit the chain elongation of [1-(14)C]20:5(n-3). This latter finding suggests that 18:4(n-3) and 20:5(n-3) might interact with the enzyme that chain elongates 20:5(n-3) to depress its ability to synthesize 22:5(n-3). Our results are most consistent with the presence of multiple chain-elongating enzymes, but a more definitive answer requires the purification of these membrane-bound proteins. In addition our results suggest that the channeling of acids between enzymes in the endoplasmic reticulum may play an important role in regulating the biosynthesis of unsaturated fatty acids.

Factors regulating the elongation of palmitic and stearic acid by rat liver microsomes.

Analysis of the rates of overall chain elongation and condensation of malonyl-CoA with palmitoyl-CoA and stearoyl-CoA as primers demonstrated that for each primer, the rate of the overall metabolic process was similar to the initial condensation. The specific activity for condensation with palmitoyl-CoA was eleven times greater than for stearoyl-CoA. The specific activities of both the beta-hydroxyacyl-CoA dehydrase and 2-trans-enoyl-CoA reductase reactions were much higher than for either condensation or chain elongation, although these rates were somewhat greater with the intermediates required in chain elongating palmitoyl-CoA than for stearoyl-CoA. Both substrates were incorporated into phospholipids at low rates and there was a time-dependent hydrolytic cleavage of the acyl-CoA primers which was partially prevented by bovine serum albumin. These findings demonstrate that there was no selective removal of either primer which could result in specific substrate depletion and an apparent reduction in the rate of condensation. These combined results firmly establish the rate-limiting nature and high degree of substrate specificity exhibited during the initial condensation step in fatty acid elongation.

Metabolism of 7,10,13,16-docosatetraenoic acid to dihomo-thromboxane, 14-hydroxy-7,10,12-nonadecatrienoic acid and hydroxy fatty acids by human platelets.

Human platelets metabolize 7,10,13,16-docosatetraenoic acid (22:4(n - 6)) into dihomo-thromboxane B2 and 14-hydroxy-7,10,12-nonadecatrienoic acid at about twenty percent of the rate they convert arachidonic acid to thromboxane B2 and 12-hydroxy-5,8,10-heptadecatrienoic acid. 14-Hydroxy-7,10,12,16-docosatetraenoic was the major metabolite produce via the lipoxygenase pathway. Several other hydroxy acids were also produced in small amounts via an indomethacin-insensitive pathway. Incubation of 20 microM arachidonic acid with various levels of 22:4(n - 6) resulted in a dose-dependent inhibition of both thromboxane B2 and 12-hydroxy-5,8,10-heptadecatrienoic acid production. Conversely, 12-hydroxy-5,8,10,14-eicosatetraenoic acid synthesis was stimulated because of substrate shunting to the lipoxygenase pathway. These results show that 22:4(n - 6) may modify platelet function both by serving as a precursor for a 22-carbon thromboxane and by suppressing the synthesis of thromboxane A2 from arachidonic acid. In addition, our results suggest that simultaneous release of 22:4(n - 6) and arachidonic acid from platelet phospholipids will result in an elevation of both 12-hydroxy-5,8,10,14-eicosatetraenoic acid levels as well as simultaneous synthesis of 14-hydroxy-7,10,12,16-docosatetraenoic acid.

Metabolism of polyunsaturated (n-3) fatty acids by monkey seminal vesicles: isolation and biosynthesis of omega-3 epoxides.

Monooxygenases of monkey seminal vesicles can metabolize arachidonic acid (20:4(n-6)) by w3-hydroxylation to 18(R)-hydroxyeicosatetraenoic acid (18(R)-HETE) and eicosapentaenoic acid (20:5(n-3)) to 17,18-dihydroxyeicosatetraenoic acid (Oliw, E.H. (1989) J. Biol. Chem. 264, 17845-17853). The present study aimed to further characterize the oxygenation of (n-3) polyunsaturated fatty acids. 14C-Labelled 22:6(n-3), 20:5(n-3), 20:4-(n-3) and 18:3(n-3) were incubated with microsomes of seminal vesicles of the cynomolgus monkey, NADPH and a cyclooxygenase inhibitor, diclofenac, and the main metabolites were identified by capillary gas chromatography-mass spectrometry. 22:6(n-3) was slowly metabolized to 19,20-dihydroxy-4,7,10,13,16-docosapentaenoic acid, while 20:5(n-3), 20:4(n-3) and 18:3(n-3) were metabolized more efficiently to the corresponding w4,w3-diols. The w3 epoxides, which were obtained from 20:5(n-3) and 18:3(n-3), were isolated in the presence of an epoxide hydrolase inhibitor, 1(2)epoxy-3,3,3-trichloropropane, and the geometry of the epoxides was determined to be 17S, 18R and 15S, 16R, respectively. While 20:5(n-3) was metabolized almost exclusively to the epoxide and diol pair of metabolites, 18:3(n-3) was metabolized not only to the w3 epoxide and the corresponding diol, but also to the w2 alcohol, 17(R)-hydroxy-9,12,15-octadecatrienoic acid. 22:6(n-3) and 5,8,11,14-eicosatetraynoic acid inhibited the biosynthesis of 18(R)-HETE from arachidonic acid (IC50 0.16 and 0.14 mM, respectively). In comparison with 20:4 or 18:3(n-3), 18:1(n-9) and 22:5(n-6) appeared to be slowly metabolized by seminal monooxygenases, while 18:2(n-6) was converted to the w3 alcohol and to smaller amounts of the w2 alcohol (4:1). Together, the results indicate that the w3-hydroxylase and w3-epoxygenase enzyme(s) metabolize 20:4(n-6) and 20:5(n-3) almost exclusively to the w3(R) alcohol and the w3(R, S) epoxide, respectively, while longer and shorter fatty acids either are poor substrates or metabolized with a lesser degree of position specificity.

The absence of an 8-desaturases in rat liver: a reevaluation of optional pathways for the metabolism of linoleic and linolenic acids.

11-[3- -14 C] Octadecenoic acid, 11,14-[1- -14C] nonadecadienoic acid, 11,14-[1- -14 C] eicosadienoic acid, 11,14-[1- -14 C] heneicosadienoic acid and 11,14,17-[3- -14 C] eicosatrienoic acid were fed and injected into tail veins of rats raised on balanced and fat-free diets. Analysis of the total liver lipids showed that 11-ated to 5,11,14-19:3; 11,14-20:2 was desaturated to 5,11,14-20:3; 11,14-21:2 was desaturated to 5,11,14-21:3 and 11,14,17-20:3 was desaturated to 5,11,14,17-20:4. In rats raised on balanced diets 11,14-19:2 and 11,14-21:2 were desaturated at very slow rates. The 11,14-20:2 and 11,14,17-20:3 were desaturated to 5,11,14-20:3 and 5,11,14,17-20:4but not as rapidly as in rats raised on fat-free diets. These findings strongly suggest that rat liver does not have a desaturase capable of introducing a double bond in the 8-position during polyunsaturated fatty acids biosynthesis. From these studies it can be concluded that linoleate is only converted to arachidonate via the following pathway: 9,12-18:2 yields 6,9,12-18:3 yields 8,11,14-20:3 yields 5,8,11,14-20:4. Linolenate is only metabolized as follows: 9,12,15-19:3 yields 6,9,12,15-18:4 yields 8,11,14,17-20:4yields 5,8,11,14,17-20:5 yields 7,10,13,16,19-22:5 yields 4,7,10,13,16,19-22:6. Optional pathways such as 9,12-18:2 yields 11,14-20:2 yields 8,11,14-20:3 yields etc. and 9,12,15-18:3 yields 11,14,17-20:3 yields 8,11,14,17-20:4 yields etc. are inoperative because 11,14-20:2 and 11,14,17-20:3 are probably not produced in vivo and if produced may well be converted to 9,12-18:2 and 9,12,15-19:3, respectively, by retroconversion.

The biosynthesis and turnover of different molecular species of rat testicular choline phosphoglycerides and triacylglycerols following intratesticular injection with (1(3)-14C)glycerol.

Rats were sacrificed 1, 4, 8, 24, 48 and 96 h after an intratesticular injection with [1(3)-14-C]glycerol. The choline phosphoglycerides and triacylglycerols were isolated and the choline phosphoglycerides were converted to diacylglycerols by treatment with phospholipase C. These diacylglycerols and the triacylglycerols were fractionated into molecular species using argentation thin-layer chromatography. At early time periods the percent of radioactivity incorporated into the oleate- and linoleate-containing diacylglycerols exceeded the molar contribution of these two fractions. The percent of radioactivity in the linoleate fraction declined rapidly at later time periods while the percent of radioactivity in the oleate fraction also declined but not as rapidly. In contrast the percent of radioactivity in the arachidonate and docosa-4,7,10,13,16-pentaenoate-containing diacylglycerols at early time periods was lower than the molar amounts of these diacylglycerols. At later time periods the percent of radioactivity in these two fractions increased. These findings suggests that oleate- and linoleate-containing choline phosphoglycerides are made largely by total synthesis while a deacylation-acylation pathway plays a major role in the biosynthesis of the highly unsaturated choline phosphoglycerides. The calculated apparent half-lifes for the oleate-, linoleate-, arachidonate- and docosa- 4,7,10,13,16-pentaenoate-containing diacylglycerols were respectively 35, 18, 41 and 46 h. The percent incorporation of [1(3)-14-C]glycerol into the various triacylglycerol fractions did not differ markedly from the molar contribution of the respective molecular fractions suggesting that the various molecular species of rat testicular triacylglycerols are all synthesized and turn over at about the same rate.

Studies to determine the role rates of chain elongation and desaturation play in regulating the unsaturated fatty acid composition of rat liver lipids.

Rat liver microsomes were used to measure the rates of chain elongation and desaturation of acids in the linoleate, oleate and palmitoleate biosynthetic pathways. These studies were designed to determine whether there is a relationship between rates of conversion and the types of unsaturated fatty acids found in rat liver lipids. In some cases rates of conversion correlate well with the types of unsaturated fatty acid found inrat liver lipids. In other cases, rates of conversion must be correlated with other controls such as competitive interactions, retroconversion, and specificities for incorporating given acids into lipids in order to explain the unsaturated fatty acid composition of rat liver lipids. The roles and interrelationships of these various metabolic processes are discussed relative to the control of polyunsaturated fatty acid biosynthesis.

Metabolism of 4,7,10,13,16-docosapentaenoic acid by human platelet cyclooxygenase and lipoxygenase.

Washed human platelets are shown to metabolize 4,7,10,13,16-docosapentaenoic acid into three major metabolites which were purified by reverse-phase HPLC. The mass spectra of the methyl ester-trimethylsilyl ether and ethyl ester-trimethylsilyl ether of compound A established it as delta 4-dihomo-thromboxane B2. Compound B was shown to be 14-hydroxy-4,7,10,12-nonadecatetraenoic acid, which is analogous to 12-hydroxy-5,8,10-heptadecatrienoic acid from arachidonic acid. Compound C was produced via an indomethacin-insensitive pathway and was identified as 14-hydroxy-4,7,10,12,16-docosapentaenoic acid. Time- and substrate-dependent studies showed that compounds A,B and C were produced approximately 10,15 and 65% of the extent to which thromboxane B2, 12-hydroxy-5,8,10-heptadecatrienoic acid and 12-hydroxy-5,8,10,14-eicosatetraenoic acid were produced, respectively, from arachidonic acid.