Degradation parameters of amaranth, barley and quinoa in alpacas fed grass hay.

This study was conducted to determine the compartment 1 (C1) characteristics of alpacas (fistulated male, 7 ± 1.5 years old, 61 ± 5 kg BW) fed grass hay (GH) supplemented with amaranth (AM), quinoa (Q) and barley (B) grains. Alpacas were provided water ad libitum while housed in metabolism crates. The GH and GH plus treatments were fed at 0700 every day. Treatment periods were for 14 days in which GH or GH plus one of the grain treatments were randomly allocated. On day 14, volatile fatty acids (VFA), pH and ammonia nitrogen (NH3 -N) were determined at 1, 3, 6, 10, 14, 18 and 24 h post-feeding. C1 degradation of each feed component was also determined with the alpacas being fed GH only and the samples incubated for 0, 2, 4, 8, 14, 24, 48 and 72 h. Dry matter (DM), neutral detergent fibre (NDF) and crude protein (CP) were determined and were divided into three categories: a = immediately soluble; b = the non-soluble but degradable; and u = non-degradable/unavailable, potential extent of degradation (PE), degradation rate (c) and effective degradation (ED). C1 passage rate was determined using acid detergent insoluble ash as a marker and was calculated to be 5.5%∙h-1. Total DM intake was highest (p < 0.05) for B and resulted in a higher (p < 0.05) CP intake. GH and AM were different in mean pH (6.81 and 6.66, respectively). B NH3 -N was greater (p < 0.05) than the other treatments. Total VFA was greatest (p < 0.05) for AM, with the greatest composition differences being a shift form acetate percentage to butyrate. DM, NDF and CP degradation was different across the treatments, where PE and ED were higher (p < 0.05) for the grain treatments. The pseudo-grains AM and Q had similar C1 degradation characteristics to B.

Isolation and characterization of a glycoprotein allergen, Art v II, from pollen of mugwort (Artemisia vulgaris L.).

A glycoprotein allergen, Art v II, was isolated from pollen of mugwort by two different isolation procedures. Art v II-A was isolated by a combination of ion-exchange chromatography on DEAE-Sepharose, affinity chromatography on Con A-Sepharose and ion-exchange chromatography on Mono P. Art v II-B was isolated by a combination of preparative IEF in Ultrodex granulated gel, affinity chromatography on Con A-Sepharose and HPLC size exclusion on Ultropac TSK G2000SW. Art v II-A and Art v II-B were shown to be antigenically identical with the allergen we have formerly denoted Ag7. The MW of Art v II A/B was determined to be 34,000-38,000 by HPLC size exclusion, and 35,000 and 20,000 by SDS-PAGE under non-reducing and reducing conditions, respectively. Art v II was found to consist of 6(7) isoforms with pI 4.10, 4.20 (major component), 4.35, 4.45, 4.55, 4.65, (4.80). The glycoprotein allergen had a protein to carbohydrate ratio of 10:1 and the carbohydrate part contained mannose (70.7%), N-acetyl-glucosamine (17.0%), glucose (7.0%) and galactose (5.3%). In R(R)IE the purified allergen bound IgE from 5 (33%) of 15 sera from patients with clinical allergy against mugwort pollen and from 13 (52%) of 25 sera from patients selected only on the basis of a RAST-class 4 against mugwort pollen.

Identification and characterization of important allergens from mugwort pollen by IEF, SDS-PAGE and immunoblotting.

Mugwort (Artemisia vulgaris L.) pollen allergens, separated by SDS-PAGE or IEF, were identified after transfer to NCM by incubation with a panel of sera from 16 patients with clinical mugwort pollen allergy, followed by [125I]anti-IgE and autoradiography. Of the at least 23 components separated by SDS-PAGE in a 15% polyacrylamide gel, at least 15 components with mol. wts 12,000-100,000 bound IgE from the panel of patient sera. A component of mol. wt 22,000 bound IgE from at least 94% of the patient sera tested and for all but three sera this component also bound the greatest quantity of IgE. Five other components with mol. wts 12,000, 17,000, 29,000, 39,000 and 42,000 bound IgE from 75-94% of the patient sera. After separation by IEF, at least 28 protein bands were detected in the pI region 3.5-7.2 and at least seven bands were found in the region 8.6-9.3. At least 11 bands in the pI range 4.2-7.3 and at least five bands in the pI region 8.5-9.2 bound IgE from the panel of patient sera. The most intense radiostaining was observed with a component having a pI of 4.35, which bound IgE from 31% of the patient sera. Immunoblotting of the SDS-PAGE and IEF gels using specific rabbit antisera and human sera against three important mugwort pollen allergens, denoted Ag 9, Ag 12 and Ag 13, was performed to determine the mol. wt and pI of these allergens which had earlier only been identified in CIE/CRIE. The results revealed that Ag 13 had a mol. wt of 61,000 and a pI of 4.35, Ag 12 had a mol. wt of 22,000 and AG 9 had pIs in the region 4.55-5.55 (six isoforms). Ag 9 did not bind IgE after SDS-PAGE and was thus not identified in the SDS-PAGE pattern, and Ag 12 failed to be detected in the NCM after transfer from IEF gels. By crossed immunoelectrofocusing, Ag 12 was found to consist of several isoforms predominantly located in the pI region 3.5-5.1. The immunoblotting analysis also revealed that the glycoprotein allergen Art v II was not detected after transfer from either SDS-PAGE or IEF gels. In conclusion, immunoblotting analysis of SDS-PAGE and IEF gels are useful methods for characterization of mugwort pollen extract, but it should be noted that some important allergens which are easily identified in CIE/CRIE may fail to be detected by these methods.

Purification of the glycoprotein allergen Ag7 from mugwort pollen by concanavalin A affinity chromatography.

Two affinity columns comprising immobilized concanavalin A (Con A), Con A-Sepharose and Con A-XP3507, were evaluated for their purifying ability for the glycoprotein allergen Ag7 from a partially purified extract of mugwort pollen. The most pronounced difference between the two columns was the nature of their nonspecific interactions; hydrophobic interactions were dominant with Con A-XP3507, whereas ionic interactions were dominant with Con A-Sepharose. Both Con A-columns were effective for purifying Ag7 with a recovery of 50% after specific elution with displacing sugars. The inclusion of 1.0 M NaCl and 20% ethylene glycol in the elution medium was useful for desorbing nonspecifically bound material, prior to specific elution of adsorbed Ag7 in the presence of the displacing sugars, alpha-methyl glucoside and alpha-methyl mannoside. The most efficient purification of Ag7 was achieved with Con A-Sepharose at room temperature rather than at 4 degrees C. Affinity chromatography with Con A-XP3507 resulted in a slightly more contaminated product (purity 54%) than with Con A-Sepharose (purity 64%).

A prospective comparison of duplex sonography vs angiography of the vertebral arteries.

A prospective blind comparison was completed between duplex sonography and angiography of the vertebral arteries. Thirty-two vertebral arteries were studied for direction of flow, degree of origin plaque or stenosis, Doppler characteristics, and vessel size. The vertebral arteries were reliably identified by imaging their course from the subclavian artery into the transverse foramina and by identifying a Doppler signal similar in waveform to the internal carotid artery. With Doppler, a 90% accuracy was obtained for direction of blood flow. Nonvisualization of origins was primarily due to vessel depth and/or tortuosity. In 12 vessel origins that were well seen with both techniques, angiography and sonography agreed in two-thirds of the cases. In four cases, origin plaque was underestimated with duplex imaging. Interestingly, no Doppler frequency or velocity elevation was identified distal to significant stenoses. One false-positive diagnosis of occlusion occurred with sonography, in which a 99% origin stenosis resulted in no detectable Doppler signal. By comparing sonography with angiography, sonography was shown to be 80% accurate in determining vertebral artery size. Our preliminary results indicate that duplex scanning is a reasonably accurate screening technique for size, patency, and direction of blood flow in the vertebral arteries. Duplex evaluation of the vertebral artery origin was limited by vessel depth, tortuosity, and calcifications.

Near infrared spectroscopy-a potentially useful method for rapid determination of fat and protein content in homogenized diets.

OBJECTIVE: To investigate the potential of near-infrared reflectance (NIR) spectroscopy as a rapid and non-destructive method to determine total fat and protein in mixed, homogenized and freeze-dried human diets. DESIGN: 29 students collected duplicate portions of their diet for four consecutive days. In addition, a detailed food diary was kept. The duplicate portions were analysed for total protein and fat content both by traditional chemical analysis (Kjeldahl and Folch methods) and through the recently developed NIR spectroscopy method. In addition, traditional computerized estimation of nutrient composition was performed. RESULTS: Plotting of the NIR-predicted fat content against the chemically analysed fat content gave a correlation coefficient of 0. 99. Plotting of the NIR-predicted protein content against the Kjeldahl-analysed protein gave a correlation coefficient of 0.81. CONCLUSION: NIR-spectroscopy seems to be able to determine fat content in mixed, homogenized diets to a high degree of accuracy. In surveys involving duplicate portion sampling this will save time and money. The prediction accuracy for protein was less convincing, but acceptable depending on the need for accurate individual data. SPONSORSHIP: Norwegian Food Research Institute, Institute for Nutrition Research at the University of Oslo and the Research Society of the Norwegian Edible Fat Producers and the food company Mills DA. European Journal of Clinical Nutrition (2000) 54, 20-23

New methods to use fish cytochrome P4501A to assess marine organic pollutants.

A new methodology has been developed to assess cytochrome P4501A expression in two South Atlantic Spanish fish, guilthead seabream (Sparus aurata) and grey mullet (Liza aurata), used as pollution bioindicators. Degenerate oligos were used to amplify by reverse transcription and PCR (RT-PCR) specific cyp1A cDNA sequences, used subsequently to design specific primers to get the full cDNA by rapid amplification of cDNA ends. A new assay has been developed to quantitate cyp1A expression by RT-PCR in an automated DNA sequencer. The effect of beta-naphthoflavone inducing biotransformation has been used to compare three distinct pollution biomarkers: EROD activity, ELISA determination of CYP1A, and 2-aminoanthracene (2-AA) activation. Immunodetection by ELISA or Western blot was inconsistent in S. aurata and L. aurata. EROD activity yielded satisfactory results; the higher induction was observed by bioactivation of 2-AA to mutagens detected with strain BA149 of Salmonella typhimurium, in agreement with the high sensitivity previously described for this biomarker. The present paper summarizes the current status of our research.

On-line prediction of chemical composition of semi-frozen ground beef by non-invasive NIR spectroscopy.

The chemical composition of industrial scale batches of frozen beef was measured on-line during grinding by near infrared (NIR) reflectance spectroscopy. The MM55E filter based non-contact NIR instrument was mounted at the outlet of a meat grinder, and the fat, moisture and protein contents determined from the average of each filter reading throughout the grinding of the batch. The filters were selected from full spectra measurements to be as insensitive to water crystallization as possible. For on-line calibration and prediction, 55 beef batches of 400-800 kg in the range of 7.66-22.91% fat, 59.36-71.48% moisture, and 17.04-20.76% protein, were ground through 4 or 13 mm hole plates. The regression results, presented as root mean square error of cross validation (RMSECV) were between 0.48 and 1.11% for fat, 0.43 and 0.97% for moisture and 0.41 and 0.47% for protein.

Prediction of sensory characteristics of beef by near-infrared spectroscopy.

Sensory hardness, tenderness and juiciness of M. Longissimus dorsi muscles from 10 beef carcasses at three ageing stages were predicted by near-infrared (NIR) spectroscopic analysis in the reflection (NIRR) and transmission modes (NIRT) during 14 days ageing at 2°C. Predicting the sensory variables hardness and tenderness from NIRR measurements using principal component regression (PCR), yielded correlation coefficients in the range 0·80-0·90. The root mean square errors of prediction for the predictions of hardness and tenderness were in the range 0·5-0·7, given in sensory assessment units. Juiciness was not well predicted. Prediction of sensory variables from NIRT measurements did not give satisfactory results. Including samples from all carcasses, cows and young bulls in the models resulted in good predictions from NIRR measurements of frozen and thawed samples. However, the best prediction results were generally obtained from separate calibrations of the samples from the bulls. The potential of NIR spectroscopy in the prediction of sensory variables in whole meat needs to be further investigated on a larger number of samples with different breeds, animals and process treatments included.

On-line, proximate analysis of ground beef directly at a meat grinder outlet.

The fat, moisture and protein contents of ground beef were determined on-line by a diffuse reflectance near infrared (NIR) spectroscopy instrument at the outlet of a meat grinder. Beef samples in the range of 6.2-21.7% fat, 59.6-72.9% moisture and 18.1-20.7% protein were studied. Calibrations from samples ground with hole diameters of 4, 8, 13 or 19 mm in the grinder plate were validated. In addition, calibrations of combinations of these samples from the different hole diameters were validated. Prediction errors, expressed as root mean square error of cross validation of the beef samples, were 0.73-1.50% for fat, 0.75-1.33% for moisture and 0.23-0.32% for protein, depending on the hole diameter of the grinder plate. Calibrations from samples ground with the smallest hole diameters gave lowest prediction errors. The present prediction error results are only slightly higher compared to reported prediction error results using conventional at- and off-line NIR instruments. It is concluded that the on-line NIR prediction results were acceptable for samples ground with grinder plates of 4, 8 or 13 mm hole diameter.

On-line NIR analysis of fat, water and protein in industrial scale ground meat batches.

Fat, water and protein contents in industrial scale meat batches were determined on-line by near infrared (NIR) reflectance spectroscopy. The NIR instrument was mounted at the outlet of a large meat grinder, and the measurements were performed in an industrial environment. Beef and pork samples, with chemical compositions of 7-26% fat, 58-75% water and 15-21% protein, were processed with hole diameters of 13mm in the grinder plate. Calibrations were made both for a combined set of beef and pork samples, and for separate sets of beef and pork samples. Validations were either done by full cross validation of the calibration set, or by bias corrected prediction of a test set. Prediction errors for the two sample sets, expressed as root mean square errors of cross validation or standard error of prediction, were in the ranges 0.82-1.49% fat, 0.94-1.33% water and 0.35-0.70% protein, depending of sample set and species of animal. The presented application is an improvement to the existing manual meat standardisation procedure, and has been implemented for regular use in a Norwegian meat manufacturing plant.

Combined effects of chilling rate, low voltage electrical stimulation and freezing on sensory properties of bovine M. longissimus dorsi.

The single and combined effects of low voltage electrical stimulation (LVES), chilling rate after slaughter, and freezing/thawing during ageing on the tenderness of bovine M. longissimus dorsi were studied. Of 27 young bulls, 14 were exposed to LVES (90 V, 32 s, 15 Hz) immediately after stunning. Four different treatments regarding chilling rate and freezing/thawing were randomly assigned to each of the 27 young bulls. Samples were aged for 7 days at 4°C and analyzed after freezing and thawing. The experimental design allowed direct comparison of different treatment effects and an assessment whether these were additive or not. As expected chilling rate after slaughter had highly significant effects on final tenderness, based on results from a trained sensory panel and Warner Bratzler (WB) shear press analysis. The effect of LVES on final tenderness was not statistically significant. The introduction of a freezing/thawing step during ageing did not significantly affect tenderness. The effects of individual treatments depended on the tenderness level and were relatively larger at high WB shear press values (7-8 kg/cm(2)) than at low values (4-5 kg/cm(2)). The results indicate that the treatment effects were not additive, but that the treatments are alternatives to tenderize beef.

Willingness to pay for diabetes drug therapy in type 2 diabetes patients: based on LEAD clinical programme results.

OBJECTIVE: The purpose of this study was to investigate the preferences of people with diabetes for liraglutide vs other glucose lowering drugs, based on outcomes of clinical trials. METHODS: Willingness to pay (WTP) for diabetes drug treatment was assessed by combining results from a recent WTP study with analysis of results from the Liraglutide Effect and Action in Diabetes (LEAD) programme. The LEAD programme included six randomised clinical trials with 3967 participants analysing efficacy and safety of liraglutide 1.2 mg (LEAD 1-6 trials), rosiglitazone (LEAD 1 trial), glimepiride (LEAD 2-3 trials), insulin glargine (LEAD 5 trial), and exenatide (LEAD 6 trial). The WTP survey used discrete choice experimental (DCE) methodology to evaluate the convenience and clinical effects of glucose lowering treatments. RESULTS: People with type 2 diabetes were prepared to pay an extra €2.64/day for liraglutide compared with rosiglitazone, an extra €1.94/day compared with glimepiride, an extra €3.36/day compared with insulin glargine, and an extra €0.81/day compared with exenatide. Weight loss was the largest component of WTP for liraglutide compared with rosiglitazone, glimepiride, and insulin glargine. Differences in the administration of the two drugs was the largest component of WTP for liraglutide (once daily anytime) compared with exenatide (twice daily with meals). A limitation of the study was that it was based on six clinical trials where liraglutide was the test drug, but each trial had a different comparator, therefore the clinical effects of liraglutide were much better documented than the comparators. CONCLUSIONS: WTP analyses of the clinical results from the LEAD programme suggested that participants with type 2 diabetes were willing to pay appreciably more for liraglutide than other glucose lowering treatments. This was driven by the relative advantage of weight loss compared with rosiglitazone, glimepiride, and insulin glargine, and administration frequency compared with exenatide.

Toxicity of medicinal plants used in traditional medicine in Northern Peru.

ETHNOPHARMACOLOGICAL RELEVANCE: The plant species reported here are traditionally used in Northern Peru for a wide range of illnesses. Most remedies are prepared as ethanol or aqueous extracts and then ingested. The aim of this study was to evaluate the potential toxicity of these extracts. MATERIALS AND METHODS: The toxicity of ethanolic and water extracts of 341 plant species was determined using a brine-shrimp assay. RESULTS: Overall 24% of the species in water extract and 76% of the species in alcoholic extract showed elevated toxicity levels to brine-shrimp. Although in most cases multiple extracts of the same species showed very similar toxicity values, in some cases the toxicity of different extracts of the same species varied from non-toxic to highly toxic. CONCLUSIONS: Traditional preparation methods take different toxicity levels in aqueous and ethanol extracts into account when choosing the appropriate solvent for the preparation of a remedy.

The occipital alpha rhythm related to the "migraine cycle" and headache burden: a blinded, controlled longitudinal study.

OBJECTIVE: Neurophysiological studies have shown a fluctuating neural dysfunction in migraine. This pathophysiological feature has not previously been investigated by quantitative electroencephalography (QEEG). The alpha rhythm is especially interesting, because it is influenced by ischemia and neuronal dysfunction within the posterior circulation area. METHODS: We investigated alpha peak frequency, variability, peak power and asymmetry in 41 migraineurs and 32 controls. Electroencephalography (EEG) was recorded on three random days and retrospectively classified as preattack, attack, postattack or interictal, based on the patient's headache diaries. We also searched for correlations between alpha rhythm parameters and disease duration, attack duration, attack frequency, pain intensity and photophobia. RESULTS: Peak frequency reduction correlated with increasing disease- and attack duration. Frequency variability increased before the attack, while peak power increased during the attack. Alpha peak width, peak frequency and peak power were similar for migraineurs and controls in the interictal period. CONCLUSION: The accumulated burden of migraine caused slight alterations in the physiology of the visual cortex. Small alpha rhythm changes were observed along the migraine cycle. SIGNIFICANCE: This is a longitudinal, controlled study. It is the first to report changes in alpha rhythm with increased migraine load, even when the QEEG is not influenced by recent or imminent attacks.

The impact of long-term haemofiltration (continuous veno-venous haemofiltration) on cell-mediated immunity during endotoxaemia.

BACKGROUND: Increased survival with high-volume continuous veno-venous haemofiltration (CVVH) has been demonstrated in critically ill patients. This may be the result of intensified blood purification or an effect on the immune system. We hypothesized that CVVH modifies the cell-mediated immunity. We investigated the effect of high-volume CVVH for 24 h on the cell-mediated immunity following endotoxin infusion. METHODS: Thirty pigs were divided into three groups. Ten pigs received 30 microg/kg of Escherichia coli endotoxin. These pigs were treated with CVVH (replacement 35 ml/kg/h) over the following 24 h. Ten pigs received the same bolus of endotoxin and ten pigs served as a control group. The adhesion molecules CD18, CD44 and CD62L and the ability to respond with an oxidative burst were measured. The number of neutrophils was counted in blood and lung tissue. The lymphoproliferative response and cytokines interleukin-6 and interleukin-10 were measured. RESULTS: The infusion of endotoxin was followed by initial granulocytopenia and, later, granulocytosis, activation of CD18 and CD62L, and increased oxidative burst. The cytokine level was increased. CVVH had no effect on the adhesion molecules or cytokine level and did not reduce the number of granulocytes in the lung significantly. CVVH, however, reduced the oxidative burst activity of neutrophils after 2 h of treatment. CONCLUSION: In the first few hours after endotoxaemia, high-volume CVVH reduced the oxidative burst activity of neutrophils. However, in the long term, CVVH was unable to modify the endotoxin-induced changes in cell-mediated immunity.

Blink reflex R2 amplitudes in cervicogenic headache, chronic tension-type headache and migraine.

Blink reflex R2 amplitude was investigated in seven patients with cervicogenic headache (CEH), 12 patients with chronic tension-type headache, 23 patients with migraine (10 with aura) and 17 headache-free controls. Standard electrical stimulation of the supraorbital nerve was applied and the response was recorded from the ipsilateral and the contralateral orbicularis oculi muscles. Low R2 amplitude was found in CEH patients compared with control subjects. Headache is unilateral in CEH and the ipsilateral and contralateral responses after stimulation on the painful side were most depressed. R2 amplitude was not significantly affected in migraine and tension headache patients. The results suggest that lower brainstem excitability is reduced in CEH. A state of hypoactivity may be present in caudal trigeminal nucleus neurons on the symptomatic side.

[Laparoscopic cholecystectomy. A new therapeutic possibility]. / Laparoskopisk kolecystektomi. Et nytt behandlingstilbud.

We present the technique of laparoscopic cholecystectomy in some detail. The method was adapted at our clinic in August 1990. Postoperative results are very promising so far. We have reason to believe that this procedure will result in fewer postoperative complications and a smoother postoperative course for the patients, enabling them to regain physical ability much earlier than after conventional biliary surgery.