Rhinopharynx irrigations and mouthwash with dissolved mupirocin in treatment of MRSA throat colonization - proof-of-concept study.

BACKGROUND: To prevent transmission of, and infection with, meticillin-resistant Staphylococcus aureus (MRSA), eradication treatment of colonized individuals is recommended. Throat colonization is a well-known risk factor for eradication failure. Staphylococcus aureus throat colonization is associated with colonization of the rhinopharynx, but in the currently recommended Danish MRSA eradication strategies, rhinopharynx colonization is not directly targeted. Rhinopharynx colonization could therefore be an important risk factor for prolonged MRSA throat carriage. AIM: To determine whether irrigation and wash of the rhinopharynx and mouth with dissolved mupirocin is a feasible and potentially efficacious supplementary strategy against treatment-resistant MRSA throat carriage. METHODS: The patient study was an open, non-blinded, trial including 20 treatment-resistant MRSA throat carriers. In the study, the patients received a supplementary treatment besides the standard treatment according to the Danish MRSA eradication strategy. The supplementary treatment consisted of rhinopharyngeal irrigation and mouth-gurgling twice a day for 14 days with a mupirocin ointment (22 g 2% ointment per litre of isotonic sterile saline solution) in a 37°C solution. FINDINGS: Eighteen patients (90%) complied with the treatment protocol and none ex-perienced any major adverse events. Out of the 18 patients who finished the study per protocol, 15 (83%) and seven (39%) patients had negative MRSA sampling results one and six months after end of treatment, respectively. CONCLUSION: This study demonstrates the feasibility and clinical potential of also targeting the rhinopharynx and oropharynx in non-systemic throat MRSA eradication strategies.

A randomized, double-blinded, placebo-controlled, parallel trial of vitamin D3 supplementation in adult patients with migraine.

BACKGROUND: Vitamin D levels have been linked to certain pain states, including migraine. This study investigated whether vitamin D supplementation would be beneficial for adult patients with migraine (ClinicalTrials.gov Identifier: NCT01695460). METHODS: A randomized, double-blinded, placebo-controlled parallel trial was conducted in migraine patients (36 women and 12 men, 18-65 years of age). A 4-week baseline period was conducted before randomization to 24 weeks of treatment. Participants were assigned to receive D3-Vitamin (n = 24, 18 women and 6 men, 100 µg/day D3-Vitamin) or placebo (n = 24, 18 women and 6 men). Migraine attacks and related symptoms were assessed by self-reported diaries. The response rate (i.e. experiencing a 50% or greater reduction in migraine frequency from baseline to week 24), change in migraine severity, and number of migraine days were recorded. Changes in migraine-related symptoms, HIT-6TM scores, and pain sensitivity tests (pressure pain threshold and temporal summation) were also evaluated. Serum levels of both 25 (OH)D and 1,25 (OH)2D were assessed from baseline to week 24. RESULTS: The number of headache days changed from 6.14 ± 3.60 in the treatment group and 5.72 ± 4.52 in the placebo group at baseline to 3.28 ± 3.24 and 4.93 ± 3.24 by the end of the trial, respectively. Migraine patients on D3-Vitamin demonstrated a significant decrease (p < .001) in migraine frequency from baseline to week 24 compared with placebo. However, migraine severity, pressure pain thresholds, or temporal summation did not show a significant change. 25(OH)D levels increased significantly for the D3-Vitamin group during the first 12 weeks of treatment. There was no significant change in 1,25(OH)2D. No side-effects were reported or noted. CONCLUSIONS: D3-Vitamin was superior to placebo in reducing migraine days in migraine patients. Larger studies are required to confirm that vitamin D3 might be one of the prophylactic options for adult patients with migraine.

Early Nutritional Follow-Up after Discharge Prevents Deterioration of ADL Functions in Malnourished, Independent, Geriatric Patients Who Live Alone - A Randomized Clinical Trial.

OBJECTIVES: To compare the effects of two nutritional follow-up interventions with regard to preventing short-term deterioration in ADL, and to compare their effects on physical function, emotional health, and health-related quality of life. DESIGN: Randomized clinical trial with two intervention groups and one control group, and a follow-up period of eight weeks. SETTING: Intervention in the participants' homes after discharge from hospital. PARTICIPANTS: Inclusion: Malnourished geriatric patients and patients at risk of malnutrition (MNA<24), aged 75 years and older, living at home and alone. Exclusion: Nursing home residents and patients with terminal illnesses or cognitive impairment. Randomization: At discharge, the patients were assigned to one of three groups: 'home visit', 'telephone consultation', or 'control' group. INTERVENTION: Individually tailored nutritional counselling of the patient and the patient's daily home carer by a clinical dietician one, two, and four weeks after discharge from hospital. The counselling was either in-person at the patients' homes, or by telephone. The control group received no follow-up after discharge. MEASUREMENTS: Primary outcome: Change in ADL (Barthel-100 score) at discharge and eight weeks later. SECONDARY OUTCOMES: Change in physical performance (handgrip strength, 30-sec. chair stand test, CAS), quality of life and depression measurements (SF-36, Depression List, Geriatric Depression Score), and Avlund mobility-tiredness score (Mob-T). RESULTS: Two-hundred and eight participants were randomized, 73 to home visits and 68 to telephone consultations. The control group comprised 67 patients. The mean age of the participants was 86.1 years. At eight weeks after discharge, 157 completed the follow-up (home visit 52, telephone consultation 51, and control group 54). The mean age of these patients was 85.8 years. More patients in the home visit group improved or maintained their ADL (96%), compared to the telephone (75%) and control groups (72%), p<0.01. No difference was detected among the groups with regard to physical measurements, health-related quality of life, and emotional health. CONCLUSION: Early nutritional follow-up after discharge, performed as home visits, prevents deterioration of ADL in malnourished, independent, geriatric patients who live alone and thereby preserves their independence.

Fish bones--a highly available calcium source for growing pigs.

In general, there is a lack of scientific documentation of nutritional value of marine by-products. The bone fraction from fish has been regarded as waste. Due to the high mineral content of fish bones, this material can be well suitable as a natural calcium source. In the present study, apparent calcium absorption of different fish bone sources was tested using growing pigs. The experimental diets consisted of boiled salmon frames, or salmon, saithe or cod bones treated with enzymes. Calcium carbonate (CaCO(3)) was used as control. The experimental diets were formulated to contain 0.7% total calcium of which the added calcium source to be tested contributed about 71% (study 1) and 86% (study 2). Except for the calcium and phosphorus sources, the animals received similar basal diets. Apparent calcium digestibility coefficient was calculated using yttrium as indicator (both studies) and was based on complete collection of faeces and urine (study 2). The experimental design was parallel and cross-over in study 1 and study 2, respectively. In study 1, piglets getting salmon bone treated with enzymes had significantly higher calcium absorption than piglets getting boiled fish bone or calcium carbonate. Therefore, in the second study only enzymatically treated fish bones were included. The higher calcium absorption from enzymatically treated salmon bone was also found in study 2, but this time not significant. Calcium from boiled salmon bones in study I, and from enzymatically treated saithe and cod bones in study II were absorbed as well as the calcium carbonate control. The results indicate that fish bones may be a useful and well absorbed calcium source. Due to the high mineral content of the bone fraction, salmon bones can be well suitable as a natural calcium and phosphorus source in, for example, food, feed or as supplement.

Plant sterols from rapeseed and tall oils: effects on lipids, fat-soluble vitamins and plant sterol concentrations.

BACKGROUND AND AIMS: Data comparing the impact of different sources of plant sterols on CVD risk factors and antioxidant levels is scarce. We evaluated the effects of plant sterols from rapeseed and tall oils on serum lipids, lipoproteins, fat-soluble vitamins and plant sterol concentrations. METHODS AND RESULTS: This was a double-blinded, randomized, crossover trial in which 59 hypercholesterolemic subjects consumed 25 g/day of margarine for 4 weeks separated by 1 week washout periods. The two experimental margarines provided 2g/day of plant sterols from rapeseed or tall oil. The control margarine had no added plant sterols. The control margarine reduced LDL cholesterol by 4.5% (95% CI 1.4, 7.6%). The tall and rapeseed sterol margarines additionally reduced LDL cholesterol by 9.0% (95% CI 5.5, 12.4%) and 8.2% (95% CI 5.2, 11.4%) and apolipoprotein B by 5.3% (95% CI 1.0, 9.6%) and 6.9% (95% CI 3.6, 10.2%), respectively. Lipid-adjusted beta-carotene concentrations were reduced by both sterol margarines (P<0.017). alpha-Tocopherol concentrations were reduced by the tall sterol compared to the rapeseed sterol margarine (P=0.001). Campesterol concentrations increased more markedly with the rapeseed sterol versus tall sterol margarine (P<0.001). The rapeseed sterol margarine increased while the tall sterol margarine decreased brassicasterol concentrations (P<0.001). CONCLUSIONS: Plant sterols from tall and rapeseed oils reduce atherogenic lipids and lipoproteins similarly. The rapeseed sterol margarine may have more favorable effects on serum alpha-tocopherol concentrations.

Monitoring the startup of a wet detention pond equipped with sand filters and sorption filters.

The startup of a wet retention pond designed for extended stormwater treatment was monitored by more than one year of continual measurement of hydraulic parameters, nutrients and quality parameters in the pond itself (pH, temperature, dissolved oxygen, turbidity). The data revealed that photosynthesis played an important role for dissolved oxygen and pH for most of the year. Another important observation was that the pond behaved more like a completely mixed reactor than like a plug flow reactor--even though the length to width ratio was as high as 4.5:1. The pond was equipped with sand filters and sorption filters whereby very good nutrient removal efficiencies were achieved.

Serum cholesterol predictive equations in product development.

The aim of the study was to incorporate trans fatty acids into predictive equations for serum cholesterol and compare their effects with the effects of the individual saturated fatty acids 12:0, 14:0 and 16:0. We have introduced trans fatty acids from partially hydrogenated soybean oil (TransV) and fish oil (TransF) into previously published equations by constrained regression analysis. Prior knowledge about the signs and ordering of existing regression coefficients were incorporated into the regression modelling by adding lower and upper bounds to the coefficients. Oleic acid (18:1) and polyunsaturated fatty acids (18:2, 18:3) were not sufficiently varied in the studies and the respective regression coefficients therefore set equal to those found by Yu et al. (Am J Clin Nutr 1995;61:1129-39). Stearic acid (18:0) considered to be neutral was not included in the equations. The regression analyses were based on results from four controlled dietary studies with a total of 95 participants and including 10 diets differing in fatty acid composition. The analyses resulted in the following equations where the change in cholesterol is expressed in mmol/L and the change in intake of fatty acids is expressed in E%: Delta Total cholesterol = 0.01 delta(12:0) + 0.12 Delta(14:0) + 0.057 delta(16:0) + 0.039 delta(TransF) + 0.031 delta(TransV)- 0.0044 delta(18:1) - 0.017 delta(18:2, 18:3) and deltaLDL cholesterol = 0.01 delta(12:0) + 0.071 delta(14:0) + 0.047 delta(16:0) + 0.043 delta(TransF) + 0.025 delta(TransV) - 0.0044 delta(18:1) - 0.017 delta(18:2, 18:3). The test set used for validation consisted of 22 data points from seven recently published dietary studies. The equation for total cholesterol showed good prediction ability with a correlation coefficient of 0.981 between observed and predicted values. The equation has been used to reformulate margarines into "trans free" products all with more favourable effects on serum cholesterol than previous products. Also a cholesterol reducing margarine has been produced. When tested against butter in an open clinical trial among subjects with mild hypercholesterolemia the observed cholesterol-lowering effect of this margarine corresponded reasonably well with the predicted (0.77 vs. 0.64 mmol/L). We conclude that the equation has practical applicability and can be used to formulate and nutritionally optimise fat products as well as to evaluate already existing products on the market.

Serum free fatty acid pattern and risk of myocardial infarction: a case-control study.

OBJECTIVES: To investigate the association between composition of serum free fatty acid (FFA) fraction and risk of a first myocardial infarction (MI). DESIGN: A case-control design. SETTING: The patients were recruited from Ullevål Hospital in Oslo and Ostfold Central Hospital in Fredrikstad and Sarpsborg, Norway. SUBJECTS: A total of 103 patients with first MI and 104 population controls, both men and postmenopausal women, age 45-75 years. RESULTS: The mean molar percentage content of docosahexaenoic (DHA), eicosapentaenoic (EPA), stearic and myristic acid in the serum FFA fraction was significantly lower in cases than in controls, whereas that of oleic and linoleic acid was higher in cases. Increased percentage content of total very long-chain omega-3 fatty acids (VLC n-3) in serum FFA was associated with decreased risk of MI. Multivariate odds ratio (OR), adjusted for age, sex, waist-hip ratio, smoking, family history of coronary heart disease (CHD) and years of education was 0.20 (95% CI 0.06-0.63) for the highest vs. lowest quartile. Also increased content of stearic acid was associated with decreased risk. Multivariate OR adjusted as above was 0.38 (95% CI 0.14-1.04) for the highest versus lowest quartile. After adjustment for oleic acid, however, the inverse linear trend was no longer significant. CONCLUSIONS: The percentage content of VLC n-3 as well as of stearic acid in serum FFA was inversely associated with risk of myocardial infarction. That of VLC n-3 may reflect diet, but additionally these free fatty acids might in some way be related to the pathogenetic process and not only reflect their content in adipose tissue.

Serum cholesterol predictive equations with special emphasis on trans and saturated fatty acids. an analysis from designed controlled studies.

The effects of dietary trans fatty acids on serum total and low density lipoprotein (LDL) cholesterol have been evaluated by incorporating trans fatty acids into predictive equations and comparing their effects with the effects of the individual saturated fatty acids 12:0, 14:0, and 16:0. Trans fatty acids from partially hydrogenated soybean oil (TRANS V) and fish oil (TRANS F) were included in previously published equations by constrained regression analysis, allowing slight adjustments of existing coefficients. Prior knowledge about the signs and ordering of the regression coefficients was explicitly incorporated into the regression modeling by adding lower and upper bounds to the coefficients. The amounts of oleic acid (18:1) and polyunsaturated fatty acids (18:2, 18:3) were not sufficiently varied in the studies, and the respective regression coefficients were therefore set equal to those found by Yu et al. [Yu, S., Derr, J., Etherton, T.D., and Kris-Etherton, P.M. (1995) Plasma Cholesterol-Predictive Equations Demonstrate That Stearic Acid Is Neutral and Monounsaturated Fatty Acids Are Hypocholesterolemic, Am. J. Clin. Nutr. 61, 1129-1139]. Stearic acid (18:0), considered to be neutral, was not included in the equations. The regression analyses were based on results from four controlled dietary studies with a total of 95 participants and including 10 diets differing in fatty acid composition and with 30-38% of energy (E%) as fat. The analyses resulted in the following equations, where the change in cholesterol is expressed in mmol/L and the change in intake of fatty acids is expressed in E%: delta Total cholesterol = 0.01 delta(12:0) + 0.12 delta(14:0) + 0.057 delta(16:0) + 0.039 delta(TRANS F) + 0.031 delta(TRANS V) - 0.0044 delta(18:1) - 0.017 delta(18:2, 18:3) and deltaLDL cholesterol = 0.01 delta(12:0) + 0.071 delta(14:0) + 0.047 delta(16:0) + 0.043 delta(TRANS F) + 0.025 delta(TRANS V) - 0.0044 delta(18:1) - 0.017 delta(18:2, 18:3). The regression analyses confirm previous findings that 14:0 is the most hypercholesterolemic fatty acid and indicate that trans fatty acids are less hypercholesterolemic than the saturated fatty acids 14:0 and 16:0. TRANS F may be slightly more hypercholesterolemic than TRANS V or there may be other hypercholesterolemic fatty acids in partially hydrogenated fish oil than those included in the equations. The test set used for validation consisted of 22 data points from seven recently published dietary studies. The equation for total cholesterol showed good prediction ability with a correlation coefficient of 0.981 between observed and predicted values. The equation has been used by the Norwegian food industry in reformulating margarines into more healthful products with reduced content of cholesterol-raising fatty acids.

Phase II clinical trial of preoperative combined chemoradiation for T3 and T4 resectable rectal cancer: preliminary results.

PURPOSE: Although preoperative chemoradiation for high-risk rectal cancer may improve survival and local recurrence rate, its adverse effects are not well defined. This prospective study evaluated the use of preoperative chemoradiation for T3 and T4 resectable rectal cancer, with special emphasis on treatment morbidity, pathologic remission rate, quality of life, and anorectal function. METHODS: Forty-two patients (30 men, 12 women) were enrolled in the study. Median distance of the distal tumor margin from the anal verge was 6.5 cm. Preoperative staging was based on digital rectal examination, endorectal ultrasound, and computed tomography. None of the patients had distant metastases. All patients had 45 Gy (1.8 Gy/day in 25 fractions) over five weeks, plus 5-fluorouracil (350 mg/m(2)/day) and leucovorin (20 mg/m(2)/day) bolus on days 1 to 5 and 29 to 33. Quality of life was assessed with the European Organization for Research and Treatment of Cancer 30-item quality-of-life questionnaire (QLQ-C30) and its colorectal cancer-specific module (QLQ-CR38) questionnaires. Objective anorectal function was assessed by anorectal manometry and pudendal nerve terminal motor latency. Surgery was performed 46 (range, 24-63) days after completion of adjuvant therapy. RESULTS: Nineteen patients (45 percent) had Grade 3 or 4 chemoradiation-induced toxic reactions. Four patients developed intercurrent distant metastases or intraperitoneal carcinomatosis at completion of chemoradiation. Thirty-eight patients underwent surgical resection: abdominoperineal resection, anterior resection, and Hartmann's procedure were performed in 55 percent, 39 percent (11 of 15 patients had a diverting stoma), and 5 percent, respectively. Major surgical complications occurred in 7 patients (18 percent) and included anastomotic leak (n = 1), pelvic abscess (n = 1), small-bowel obstruction (n = 3), and wound breakdown (n = 2). Final pathology was Stage 0 (no residual disease), I, II, and III in 6 (16 percent), 7 (18 percent), 9 (24 percent), and 16 (42 percent) patients, respectively. There was a deterioration, after chemoradiation and surgery, in the quality of life on all subscales assessed, with physical, role, and social function being most severely affected. The symptoms most adversely affected were micturition, defecation, and gastrointestinal problems. Body image and sexual enjoyment deteriorated in both men and women. Chemoradiation alone led to prolonged pudendal nerve terminal motor latency in 57 percent of 7 patients assessed. CONCLUSION: Preliminary results have identified defined costs with preoperative chemoradiation, which included treatment-induced toxicity, a high stoma rate, and adverse effects on quality of life and anorectal function.

Partially hydrogenated soybean oil reduces postprandial t-PA activity compared with palm oil.

The effects of dietary trans fatty acids on fasting and diurnal variation in hemostatic variables are not known. This study compares the effects of three diets with three different margarines, one based on palm oil (PALM-diet), one based on partially hydrogenated soybean oil (PHSO, TRANS-diet) and one with a high content of polyunsaturated fatty acids (PUFA-diet) on diurnal postprandial hemostatic variables. A strictly controlled dietary Latin square study was performed and nine young female participants consumed each of the diets for 17 days in a random order. The sum of the cholesterol-increasing fatty acids (C12:0, C14:0, C16:0) was 36.3% of total fatty acids in the PALM-diet, the same as the sum of saturated-(C12:0, C14:0, C16:0) (12.5%) and trans fatty acids (23.1%) in the TRANS-diet. The sum of C12:0, C14:0 and C16:0 was 20.7% in the PUFA-diet. The amount of fat made up 30-31% of energy in all diets. Nine participants completed the study. The diurnal postprandial state level of tissue plasminogen activator (t-PA) activity was significantly decreased on the TRANS-diet compared with the PALM-diet. t-PA activity was also decreased on the PUFA-diet compared with PALM-diet but the difference was below statistical significance (P=0.07, Bonferonni adjusted). There were no significant differences in either fasting levels or in circadian variation of t-PA antigen, PAI-1 activity, PAI-antigen, factor VII coagulant activity or fibrinogen between the three diets. Our results indicate that dietary trans fatty acids from PHSO has an unfavourable effect on postprandial t-PA activity and thus possibly on the fibrinolytic system compared with palm oil.

Carica papaya glutamine cyclotransferase belongs to a novel plant enzyme subfamily: cloning and characterization of the recombinant enzyme.

A full-length cDNA encoding Carica papaya glutamine cyclotransferase was cloned by RT-PCR on the basis of results from amino acid sequencing of tryptic fragments of the native enzyme. The cDNA of 1036 nucleotides encodes a typical 22-residue signal peptide and a mature protein of 266 residues with a calculated molecular mass of 30,923 Da. Five plant ESTs encoding putative QCs highly homologous to PQC were identified and the numbers and locations of cysteines and N-glycosylation sites are conserved. The plant QC amino acid sequences are very different from the known mammalian QC sequences and no clear homology was observed. The PQC cDNA was expressed in Escherichia coli as either His-tagged PQC, with three different signal peptides and in fusions with thioredoxin, glutathione S-transferase, and (pre-) maltose-binding protein. In all cases, the expressed protein was either undetectable or insoluble. Expression in Pichia pastoris of PQC fused to the alpha-factor leader resulted in low levels of PQC activity. Extracellular expression of PQC in the insect cell/baculovirus system was successful and 15-50 mg/liter of active PQCs with three different secretion signals was expressed and purified. Further, PQC N-terminally fused to a combined secretion signal/His-tag peptide was correctly processed by the host signal peptidase and the His-tag could subsequently be removed with dipeptidyl peptidase I. The expressed products were characterized by activity assays, SDS-PAGE, N-terminal amino acid sequencing, MALDI-TOF mass spectroscopy, and peptide mass fingerprint analysis.

Adipose tissue fatty acids and risk of myocardial infarction--a case-control study.

OBJECTIVES: To study the association between content in adipose tissue of very long-chain n-3 fatty acids, trans fatty acids, linoleic acid and alpha-linolenic acid and risk of a first myocardial infarction. DESIGN AND SUBJECTS: A case-control design among 100 patients and 98 population controls both men and postmenopausal women, age 45-75 y. Adipose tissue fatty acids were determined by gas-liquid chromatography. Intake data were obtained through interview using a validated food frequency questionnaire. RESULTS: Dietary intake and adipose tissue content of the fatty acids studied correlated significantly. Adipose tissue contents of eicosapentaenoic acid (20:5n-3), docosapentaenoic acid (22:5n-3) and docosahexaenoic acid (22:6n-3) were significantly lower while those of trans fatty acids, linoleic and alpha-linolenic acid were significantly higher in patients than in controls. Age and sex adjusted odds ratios (OR) were significantly reduced with increasing quintiles of very long-chain n-3 fatty acids, thus the OR in the fifth compared to the first quintile was 0.23 (95% CI 0.08-0.70). After further adjustment for waist-to-hip ratio, smoking, family history of CHD and content of trans fatty acids, the OR in the highest quintile was 0.17 (95% CI 0.04-0.76) and the P for trend 0. 016. Age and sex adjusted OR was increased in the fifth compared to the first quintile of trans fatty acids (OR 2.81, 95% CI 1.16-6.84), linoleic acid (OR 2.10, 95% CI 0.87-5.07) and alpha-linolenic acid (OR 1.96, 95% CI 0.83-4.61), and P for trend was 0.002, 0.005 and 0. 020, respectively. The trends remained significant after adjustment for waist-to-hip ratio, smoking, and family history of coronary heart disease. Trans fatty acids, linoleic acid and alpha-linolenic acid in adipose tissue were strongly correlated, indicating a common source, most likely margarine. When each of these fatty acid species were adjusted for the two others the trends were no longer significant. CONCLUSION: Intake of very long-chain n-3 fatty acids as reflected in adipose tissue content is inversely associated with risk of myocardial infarction. Trans fatty acids, linoleic and alpha-linolenic acid were intercorrelated and associated with increased risk. It is suggested that the increased risk may be connected to trans fatty acids or to some other factor associated with margarine consumption. European Journal of Clinical Nutrition 54, 618-625.

Interaction of pig kidney and lentil seedling copper-containing amine oxidases with guanidinium compounds.

The effect of guanidinium compounds on the catalytic mechanism of pig kidney and lentil seedling amine oxidases has been investigated by polarographic techniques and spectroscopy. Guanidine does not inhibit the lentil enzyme and is a weak inhibitor for pig kidney amine oxidase (Ki=1 mM), whereas aminoguanidine is an irreversible inhibitor of both enzymes, with a Ki value of 10(-6) M. 1,4-Diguanidino butane (arcaine) is a competitive inhibitor for both pig and lentil amine oxidases. Amiloride is a competitive inhibitor for pig enzyme, but upon prolonged incubation with this drug the enzyme gradually loses its activity in an irreversible manner.

Effect of metal substitution in copper amine oxidase from lentil seedlings.

The reaction with substrates and carbonyl reagents of native lentil Cu-amine oxidase and its modified forms, i.e. Cu-fully-depleted, Cu-half-reconstituted, Cu-fully-reconstituted, Co-substituted, Ni-substituted and Zn-substituted, has been studied. Upon removal of only one of the two Cu ions, the enzyme loses 50% of its enzymatic activity. Using several substrates, Co-substituted lentil amine oxidase is shown to be active but the k(c) value is different from that of native or Cu-fully-reconstituted enzyme, while K(m) is similar. On the other hand, the Ni- and Zn-substituted forms are catalytically inactive. Enzymatic activity measurements and optical spectroscopy show that only in the Co-substituted enzyme is the organic cofactor 6-hydroxydopa quinone reactive and the enzyme catalytically competent, although less efficient. The Co-substituted amine oxidase does not form the semiquinone radical as an intermediate of the catalytic reaction. While devoid or reduced of catalytic activity, all the enzyme preparations are still able to oxidise two moles of substrate and to release two moles of aldehyde per mole of dimeric enzyme. The results obtained show that although Co-substituted amine oxidase is catalytically competent, copper is essential for the catalytic mechanism.

CuI-semiquinone radical species in plant copper-amine oxidases.

The intermediate CuI-semiquinone radical species in the catalytic mechanism of copper-amine oxidase from Lens esculenta and Pisum sativum seedlings has been studied by optical, Raman resonance and ESR spectroscopies and by stopped-flow and temperature-jump measurements. Treatment of highly purified enzyme preparations with good, poor or suicide substrates, under anaerobic and aerobic conditions, at different pH values and temperatures, makes it possible to generate, detect and characterize this free radical intermediate.

Molecular modeling and preclinical evaluation of the humanized NR-LU-13 antibody.

A mouse-human chimeric monoclonal antibody (chNR-LU-13), specific for the EGP40 pancarcinoma antigen, was humanized through three-dimensional molecular modeling. Humanization of the chNR-LU-13 antibody is expected to enhance its use for patients undergoing immunotherapy. On the basis of the observed amino acid sequence identity, chNR-LU-13 complementary determining regions (CDRs) of the V(L) and V(H) regions were grafted onto the human anti-DNA-associated idiotype immunoglobulin clone, R3.5H5G'CL. Ten amino acids residues within the humanized framework were back-mutated to their corresponding chNR-LU-13 sequence, because they were predicted to disrupt the canonical classification of the CDRs or were within 5 A of a CDR. Synthesis of the V(L) and V(H) regions was accomplished by recursive PCR, and the dual-chain expression vector p451.C4 was positioned under control of the CMV(P+E). We observed by competitive ELISA that the recombinant humanized NR-LU-13 (huNR-LU-13) IgG1 antibody exhibited an indistinguishable immunoreactivity profile when compared with the murine monoclonal antibody (muNR-LU-10). The huNR-LU-13 antibody was effective in mediating both antibody-dependent cellular cytotoxicity and complement-mediated cytotoxicity when assayed against either the breast carcinoma cell line, MCF-7, or the colon adenocarcinoma cell line, SW1222. Biodistribution studies using i.v. coinjected 131I-muNR-LU-10 and 125I-huNR-LU-13 confirmed that the huNR-LU-13 specifically targets to the tumor in athymic BALB/c mice bearing the SW1222 human tumor xenograft. Humanization of the chNR-LU-13 antibody is expected to eliminate an undesired human antimouse antibody response, allowing for repeated i.v. administration into humans.

[Trans-fatty acids and health]. / Transfettsyrer og helse.

Trans fatty acids are formed by partial hydrogenation of vegetable and marine oils or by bacterial activity in the rumen of ruminants. Main dietary sources are margarine, meat, milk fat and bakery products. Unsaturated fatty acids in the trans form have a more straight structure than their cis counterparts. They therefore have properties more like saturated fatty acids. Trans fatty acids may compete with essential fatty acids for elongating and desaturating enzymes and thereby interfere in the formation of eicosanoids. Trans fatty acids in the diet will increase LDL-cholesterol but to a lesser degree than the saturated fatty acids C12-C16. They also decrease HDL-cholesterol and increase Lp(a). By these unfavorable effects on blood lipids it may be expected that they will increase the risk of coronary heart disease. This has been confirmed in some, but not all, epidemiological studies. Provided the diet contains sufficient amounts of essential fatty acids there are no strong indications that trans fatty acids may have other unfavorable effects on health. Over time the intake of trans fatty acids has decreased considerably in Norway (from a mean of about 15 g/day in 1958 to about 4 g/day actually). The health effects of such an intake must be considered minor compared to an eight to ten fold higher intake of saturated fatty acids.

Replacement of partially hydrogenated soybean oil by palm oil in margarine without unfavorable effects on serum lipoproteins.

We have compared the effects of three different margarines, one based on palm oil (PALM-margarine), one based on partially hydrogenated soybean oil (TRANS-margarine) and one with a high content of polyunsaturated fatty acids (PUFA-margarine), on serum lipids in 27 young women. The main purpose of the study was to test if replacement of trans fatty acids in margarine by palmitic acid results in unfavorable effects on serum lipids. The sum of saturated fatty acids (12:0, 14:0, 16:0) was 36.3% of total fatty acids in the PALM-diet, the same as the sum of saturated (12:0, 14:0, 16:0) (12.5%) and trans (23.1%) fatty acids in the TRANS-diet. This sum was 20.7% in the PUFA-diet. The content of oleic acid was 37.9, 35.2, and 38.6%, respectively, in the three diets, whereas linoleic acid amounted to 16, 13.5, and 27.3%, respectively. Total fat provided 30-31% and the test margarines 26% of total energy in all three diets. The subjects consumed each of the diets for 17 d in a Latin-square crossover design. There were no significant differences in total cholesterol, low density lipoprotein (LDL)-cholesterol and apolipoprotein B (apoB) between the TRANS- and the PALM-diets. High density lipoprotein (HDL)-cholesterol and apoA-1 were significantly higher on the PALM-diet compared to the TRANS-diet whereas the ratio of LDL-cholesterol to HDL-cholesterol was lower, although not significantly (P = 0.077) on the PALM-diet. Total cholesterol, LDL-cholesterol, and apoB were significantly lower on the PUFA-diet compared to the two other diets. HDL-cholesterol was not different on the PALM- and the PUFA-diets but it was significantly lower on the TRANS-diet compared to the PUFA diet. Compared to the PUFA-diet the ratio of LDL- to HDL-cholesterol was higher on both the PALM- and the TRANS-diets whereas apoA-1 was not different. Triglycerides and lipoprotein (a) were not significantly different among the three diets. We concluded that nutritionally, palmitic acid from palm oil may be a reasonable alternative to trans fatty acids from partially hydrogenated soybean oil in margarine if the aim is to avoid trans fatty acids. A palm oil-based margarine is, however, less favorable than one based on a more polyunsaturated vegetable oil.