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Introduction:There is a hierarchical order in tendon ranging from the collagen molecule to the whole tendon. We have developed a method using the atomic force microscope (AFM) together with a novel data acquisition technique to quantify the sub-failure mechanical properties of individual collagen fibrils. Here we use this method to investigate how the saline environment affects tensile force transmission.Methods:Human patellar tendon tissue was obtained from a healthy young male. Six fascicles and six fibrils were mechanically tested sub-failure to determine modulus and relative energy dissipation in PBS solutions of 0.02, 0.15 and 1 M concentration as well as two HEPES buffers containing NaCl or NaCl+CaCl2. Mechanical testing was performed to 4% strain.View Large Image | View Hi-Res Image | Download PowerPoint SlideResults:The tensile properties of collagen fibrils are highly resistant to changes in their saline environment. The only consistent response to environment composition was a minor (

Fourteen patients suffering from familial hypercholesterolemia (type IIa) participated in a double-blind, placebo-controlled trial that evaluated the effects of fish oil ethyl ester (K-85, 5.7 g/day) or a hydroxymethylglutaryl coenzyme A reductase inhibitor (lovastatin, 40 mg/day) alone or in combination on lipid metabolism and bleeding time at rest and after standardized exercise. Lovastatin treatment reduced total cholesterol (-27%), low density lipoprotein cholesterol (-37%), and triglycerides (-18%), whereas high density lipoprotein cholesterol increased significantly (14%). K-85 affected total (-4%), low density lipoprotein (-9%), and high density lipoprotein (+7%) cholesterol insignificantly, whereas the triglyceride level decreased by 24% (p < 0.001). The combined regimen caused an additive decrease in the triglyceride level (41%), which differed significantly (p < 0.01) from that gained by lovastatin alone. Under basal conditions the bleeding time was not influenced by the different interventions. Standardized exercise shortened the bleeding time by 19% (p < 0.001) and 16% (p < 0.001) before intervention and after lovastatin treatment, respectively. After K-85 alone or in combination with lovastatin, the exercise-induced shortening of the bleeding time was totally inhibited, which may reflect a favorable influence of fish oil on the platelet-vessel wall interaction in these high-risk patients.

A sex difference in the bleeding time has been reported. To investigate the effects of the different sex hormones, the bleeding time (BT) and hemostatic factors related to it were measured in 209 healthy women: 50 pregnant, 113 menstruating and 46 post-menopausal. The serum levels of estrogens, progesterone and testosterone were distributed as expected for the different groups. There were no significant differences in the BT between any of the groups. No correlations between the BT and female or male sex hormones were found. The findings indicate that although sex hormone mediated effects on platelet/endothelial cell interactions may exist, they do not become evident within the physiological variations in the BT of healthy women.

Conclusive insight into the microscopic principles that govern the strength of tendon and related connective tissues is lacking and the importance of collagen cross-linking has not been firmly established. The combined application of whole-tissue mechanical testing and atomic force spectroscopy allowed for a detailed characterization of the effect of cross-linking in rat-tail tendon. The cross-link inducing agent glutaraldehyde augmented the tensile strength of tendon fascicles. Stress at failure increased from approximately 8 MPa to approximately 39 MPa. The mechanical effects of glutaraldehyde at the tendon fibril level were examined by atomic force microscopy. Peak forces increased from approximately 1379 to approximately 2622 pN while an extended Hertz fit of force-indentation data showed a approximately 24 fold increase in Young's modulus on indentation. The effect of glutaraldehyde cross-linking on the tensile properties of a single collagen fibril was investigated by a novel methodology based on atomic force spectroscopy. The Young's modulus of a secluded fibril increased from approximately 407 MPa to approximately 1.1 GPa with glutaraldehyde treatment. Collectively, the findings indicate that cross-linking at the level of the collagen fibril is of key importance for the mechanical strength of tendon tissue. However, when comparing the effects at the level of the tendon fascicle and fibril, respectively, further questions are prompted regarding the pathways of force through the tendon microstructure as fibril strength seems to surpass that of the tendon fascicle.

Continuous recruitment and inappropriate activity of platelets in the airways may contribute to airway remodeling, a characteristic feature of inflammatory airway diseases that includes increased proliferation of the smooth muscle. The aim of the present investigation was to examine the effect of platelets on proliferation of airway smooth muscle cells (ASMC) in culture and to determine the possible role of 5-lipoxygenase (5-LOX) and reactive oxygen species (ROS) in this context. ASMC obtained from guinea pigs were cultured and co-incubated with washed platelets for 24 hours. Thereafter, the proliferation was measured with the MTS-assay, the results were also verified by using thymidine incorporation, DNA measurements and manual counting. The interaction between platelets and ASMC was visualised with fluorescence microscopy. We found that platelets bind to the ASMC and the presence of platelets caused a significant dose-dependent increase in ASMC proliferation. Co-incubation of ASMC with platelets also increased ROS-production, detected by the fluorescent probe DCFDA. Furthermore, the platelet-induced proliferation was reduced in the presence of the NADPH-oxidase inhibitors DPI and apocynin. A possible role of 5-LOX in platelet-induced proliferation and ROS-generation was evaluated by using the 5-LOX inhibitor AA-861 and the PLA2-inhibitor ATK. The results showed that inhibition of these enzymes significantly reduced the platelet-induced proliferation. Moreover, Western blot analysis revealed that the ASMC but not the platelets express 5-LOX. In addition, our experiments revealed that the presence of AA-861 and ATK significantly inhibited the ROS-production generated upon coincubation of platelets and ASMC. In conclusion, we show that platelets have a marked capacity to induce ASMC proliferation. Furthermore, our study indicates that the interaction between platelets and ASMC leads to activation of 5-LOX in the ASMC followed by an increased ROS-production, events resulting in enhanced ASMC proliferation. The new findings are of importance in understanding possible mechanisms contributing to airway remodeling. Original publication: Ann-Charlotte B. Svensson Holm, Torbjörn Bengtsson, Magnus Grenegård and Eva G. Lindström, Platelets stimulate airway smooth muscle cell proliferation through mechanisms involving 5-lipoxygenase and reactive oxygen species, 2008, Platelets, (19), 7, 528-536.http://dx.doi.org/10.1080/09537100802320300. Copyright © Taylor & Francis Group, an informa business

In 1987 we noticed excess adipose tissue in a patient with arm lymphedema and later, objective studies confirmed this clinical finding in patients with non-pitting arm lymphedema following breast cancer. A prospective study was begun in 1993, and its long-term results (15 years) shows overall complete reduction of the excess volume in patients with non-pitting arm lymphedema and that adipose tissue dominates the excess volume. Encouraged by these results we operated on a patient with primary and secondary elephantiasis of the leg. The edema was first transferred from a pitting to a non-pitting state by controlled compression therapy. Then liposuction was performed to remove the remaining excess adipose tissue, and complete reduction was finally achieved. The patient wears compression garments continuously and during the 11 years of followup, no recurrence has occurred. This paper explains our philosophical approach: a pitting lymphedema first should be treated conservatively to remove excess fluid, then liposuction can be performed to remove remaining excess volume bothersome to the patient.

Audiological testing, interviews and exposure measurements were used to collect data on the health effects of styrene exposures in 313 workers from fiberglass and metal-product manufacturing plants and a mail terminal. The audiological test battery included pure-tone audiometry, distortion product otoacoustic emissions (DPOAE), psychoacoustic modulation transfer function, interrupted speech, speech recognition in noise and cortical response audiometry (CRA). Workers exposed to noise and styrene had significantly poorer pure-tone thresholds in the high-frequency range (3 to 8 kHz) than the controls, noise-exposed workers and those listed in a Swedish age-specific database. Even though abnormalities were noted on DPOAE and CRA testing, the interrupted speech and speech recognition in noise tests were the more sensitive tests for styrene effects. Further research is needed on the underlying mechanisms to understand the effects of styrene and on audiological test batteries to detect changes in populations exposed to solvents.

This paper gives an overview of the ongoing activities under the Antenna Measurement activity of the Antenna Centre of Excellence (ACE) network within the EU 6th framework research program. In particular, in this work an attempt is made to establish a common uncertainty estimation criteria in spherical near field and far field antenna measurement systems. The results from this activity are important instruments to verify the measurements accuracies for antenna measurement ranges as well as to investigate and evaluate possible improvements in measurement set-ups and procedures. These results will be used in the facility comparison campaigns in order to calculate a reference pattern for each of the high accuracy reference antennas (VAST 12, SATIMO SH800 and SATIMO SH2000) measured during the last 4 years by different institutions in Europe and US. (6 pages)

Different teat-cleaning methods were evaluated to determine their effect on the presence of spores from anaerobic bacterial spore-formers in the milk. Artificial contamination was used to achieve uniform contamination of teats to reduce the number of cows and samples needed in the experiments and still obtain adequate power to detect differences among tested methods. Teats were contaminated experimentally with a large amount of Clostridium tyrobutyricum spores in a manure-water slurry. Various types of dry and moistened towels and different combinations of methods using soap or 2 types of towels, together with cleaning times of 10 or 20 s, were compared in 2 Latin square-designed experiments with 7 cows, 7 treatments, and 4 replications in each experiment. In comparison with control (no cleaning and no forestripping), cleaning teats with dry paper towels for 10 s reduced concentration of spores in milk by 45 to 50%. A 50 to 74% reduction was achieved using different types of moist towels for 10 s. Methods using 2 towels, soap, or a longer cleaning time reduced bacterial contamination by 85 to 91%. The most effective methods in reducing milk spore content (96% reduction) were use of a moist washable towel with or without soap followed by drying with a dry paper towel, for a total time of 20 s per cow. One of the best cleaning methods was studied in an additional experiment to determine the effect of different teat contamination mixtures. The Latin square-designed experiment with 8 cows, 8 treatments, and 2 replications showed that cleaning was independent of the tested contamination matrix (manure, soil, or sawdust), type of spores (Cl. tyrobutyricum and Bacillus cereus), or degree of contamination (manure or extra manure).

Concepts such as modularity and heterogeneity are becoming important for a growing number of applications that use massively parallel computer architectures. Application areas which seem to require these concepts appear in real world computing and action oriented systems. In many instances the current offerings of high pevormance, parallel, general purpose computers are not well suited to these applications since they do not address issues like real-time, time determinism, heterogeneous communication, physical size, power consumption, etc. These issues are important in special systems that can be viewed as aon orming to general purpose markets. The differences in needs will be explored by looking into two examples of modular and heterogeneous systems: high pe ion systems and action oriented me

The beliefs and attitudes regarding the risk of hearing loss and their impact on hearing protector use were investigated among Swedish workers. A questionnaire, developed by the US National Institute for Occupational Safety and Health (NIOSH), was used. The study objective was to assess workers' attitudes towards using hearing protection devices (HPDs) and to enhance the ability of workers to protect themselves from occupational hearing loss. Ninety-five per cent of the respondents were aware that loud noise could damage their hearing, 90% considered that a hearing loss would be a serious problem, and 85% believed that HPDs could protect their hearing. However, lower percentages of workers always used the HPDs when they were noise-exposed. Fifty-five per cent of the workers indicated that they could not hear warning signals when using HPDs, and 45% of the workers indicated that they considered HPDs to be uncomfortable. These issues must be addressed to make HPD use more effective.

Tissue-factor-pathway inhibitor is the principal regulator of tissue factor-induced coagulation. Heparin treatment mobilizes TFPI into the circulation and contributes to the anticoagulant effects of heparins. Previous studies have demonstrated a selective depletion of intravascular TFPI by unfractionated heparin (UFH) but not by low-molecular-weight heparin (LMWH). In this study we sought to investigate the time- and dose-dependent relationships between release of TFPI and lipoprotein lipase (LPL) in respons to UFH and LMWH and to investigate whether the selective depletion of TFPI by UFH but not by LMWH is related to differential urinary excretion of TFPI. Eight healthy males participated in an open crossover study in which participants were assigned to receive (1) continuous infusion of unfractionated heparin (UFH, 450 IU/kg/24 hr); (2) subcutaneous dalteparin, 100 IU/kg given twice at a 12-hr interval; (3) subcutaneous dalteparin, 200 IU/kg given once; or (4) saline-solution infusion. Similar dose-dependent mobilization of TFPI and lipoprotein lipase (LPL), another glucosaminoglycan (GAG)-anchored protein of the endothelial membrane, was observed after both subcutaneous and intravenous administration of heparins. However, UFH induced a more efficient release of both TFPI and LPL into plasma than did LMWH at equivalent anti-Xa levels, indicating molecular-weight dependence of the release reactions. However, LPL reached peak levels faster and was more rapidly cleared from the circulation than was TFPI, regardless of the treatment modality. Only trace amounts of TFPI were detected in the urine in a native form (38 kD). UFH and LMWH treatment reduced renal clearance of TFPI compared with the control regimen. Our findings suggest that displacement of TFPI from the endothelial-surface GAG is the main mechanism for TFPI release during heparin treatment in vivo and that differential urinary excretion of TFPI is not the explanation for selective depletion of TFPI during UFH treatment.

Allergic rhinitis is a common health problem usually treated with drug therapy. Some patients experience side effects of drug therapy while others fear the use of drugs. Acupuncture is an interesting alternative to traditional treatment. The few studies evaluating acupuncture indicate a possible clinical effect on allergic rhinitis. This study compared active versus sham acupuncture in 40 consecutive patients with a history of allergic rhinitis and a positive skin test. Patients were randomized and assessed prior to treatment and then reassessed after 12 months. Improvements in symptoms using visual analogue scales, reduction in skin test reactions and levels of specific immunoglobin E (IgE) were used to compare the effect of treatment. For one allergen, mugwort, a greater reduction in levels of specific IgE ( p =0.019, 0.039) and skin test reaction ( p =0.004) was seen in the group receiving active acupuncture compared to the group receiving sham acupuncture. However, this finding might be an artifact. No differences in clinical symptoms were seen between active versus sham acupuncture, thus the conclusion being that the effect of acupuncture on allergic rhinitis should be further evaluated in larger randomized studies.

Objective. To evaluate if magnetic resonance imaging (MRI) is superior to conventional radiography for detection of erosions in the fifth metatarsophalangeal (MTP5) joint. Methods. Within one year from the onset of rheumatoid arthritis (RA) (baseline), one and three years thereafter MRI and conventional radiographs of the MTP5 joint were performed in 23 patients. Results. MRI revealed erosions in 10 patients at baseline, in 15 after one year and in 15 patients after 3 years. On conventional radiography, there were erosions in 10 patients at baseline, 16 after one year as well as after 3 years. The agreement between the two imaging methods was fair to good at baseline and after one and three years (kappa 0,65, 0,51 and 0,51 respectively). The number of patients with clinical evidence of synovitis decreased considerably over time although the number of patients with MRI-synovitis was unchanged and the number of patients with erosions increased. Conclusions. MRI was not superior to conventional radiography in detecting erosions in MTP5 joints in patients with early RA. Most erosions developed during the first year of observation. Synovitis on MRI may be a marker of future development of erosions in the MTP5 joint. (Less)

To study the outcome in clinical practice of first DMARD and/or corticosteroid (CS) treatment in patients with recent onset rheumatoid arthritis (RA).245 patients with active RA, not previously treated with DMARDs or CS, were randomised to one of two treatment groups, T1 = 7.5-15 mg of prednisolone (PRE) daily for one to three months followed, if needed, by methotrexate (MTX) in a weekly dose of 5-15 mg in addition to the lowest possible dose of PRE or T2 = sulfasalazine (SAL), supplemented with lowest possible CS dose if needed.The EULAR individual response criteria were applied and remission was defined as a final DAS282.6. Function was assessed by the HAQ and radiographic progression by Larsen scores. A patient who managed to remain on the allocated treatment for two years was described as a "completer".After 2 years of treatment, 70% of the patients in T1 and 63% in T2 were responders (30% and 33% "good responders", respectively). In T1 29% and in T2 19% were in remission. There was a significant functional improvement in both groups but radiographic progression occurred. The mean decrease in HAQ and increase in the Larsen score were similar in the two groups. One-third of the patients were non-completers, 19% from T1 and 47% from T2. Non-completers had, compared with completers, a significantly lower rate of individual response and remission. Completers and non-completers had similar functional improvement and similar radiological progression.Individual response and remission was reduced in patients who did not complete their first DMARD/CS treatment option. Treatment failures were significantly more frequent in the sulfasalazine plus optional CS than in the CS plus optional methotrexate treatment group.

This paper addresses two related problem areas with regard to autonomous systems, namely (i) appropriate architectures of the embedded computing systems, and (ii) appropriate system development strategies and tools. Adaptivity and learning are identified as important characteristics of autonomous systems. A modular architecture capable of executing multiple-neural-network systems with real-time learning and control abilities is described. This architecture is based on distributed nodes of massively parallel processing elements communicating by means of a scalable high bandwidth medium. The nodes and the medium operate in a time-deterministic manner. Further, it is argued that autonomous systems must be developed through a process of system alteration-change-orientation rather than the phase-oriented specify-design-implement-and-test model which is common today. A tool which, in cooperation with the system architecture, supports this paradigm is described. Required functions of the development tool are, e.g., inspection and visualization of data, change of software and hardware functions, and ability to cope with changes in the mechanical structure, including sensors and actuators, all while the autonomous system is in action. A graphical, interactive design environment that meets these demands is outlined. >

Considers DP (distributed/parallel) Cy-Clone, which is a further development of previous work in the area of designing highly predictable real-time systems. DP Cy-Clone systems contain only architectures that have well controlled time behavior describable in terms of time increments (dT). The issues related to designing time-critical distributed/parallel real-time systems and the motivation for the properties of DP Cy-Clone are presented first. The generic architecture of a node and a distributed network of nodes is presented, followed by the introduction of one possible approach to a bus-based concrete architecture. Means of network expansion via connecting multiple clusters are presented. Finally, some potential areas of application are described. >

The application of artificial neural networks (ANN) in real-time embedded systems demands high performance computers. Miniaturized massively parallel architectures are suitable computation platforms for this task. An important question which arises is how to establish an effective mapping from ANN algorithms to hardware. In this paper, we demonstrate how an effective mapping can be achieved with our programming environment in close combination with an optimized architecture design targeted for neuro-computing. >

Slot antennas have been widely used for military and defense applications for decades. This antenna type offers a compact and attractive solution in many cases. The use of the waveguide technique gives a low loss design and will often result in a well defined performance. Slot models and design tools are mature and present an opportunity to do most of the design work theoretically without tedious measurements and experimental efforts. The development and refinement of models for all kinds of slots has been going on since the 1940s and the literature still contains many examples of applications for slotted waveguide arrays. The slotted waveguide antenna is therefore an attractive alternative also for commercial applications. This is especially true for high frequency applications such as radio links where several frequency bands are allocated. Radar antennas and point-point antennas are mentioned in particular and radiation patterns discussed.

Ergometer cycling performance as well as acute exercise-induced changes in the metabolism of energy-intermediates and glutathione (GSH) were investigated in skeletal muscle (SM) of 15 healthy young male subjects (VO(2max) approximately 54.7 mL kg(-1) min(-1), age approximately 25 years), before and after 3 days of controlled 'ìoverload-training' in combination with either high (62% of energy intake) or low (26% of energy intake) dietary intake of carbohydrates. The intake of a carbohydrate-rich diet clearly reduced the depletion of SM glycogen following the short-term training period, paralleled with a positive effect on the endurance performance, but not on high-intensity work-performance. An 'delayed over-reaching effect', defined as impaired work-performance, was observed after 2.5 days of recovery from the short-term training period, irrespective of the carbohydrate content of the diet and basal glycogen level in SM. Taken together, the main and novel findings of present investigation are: (1) an acute decrease of reduced GSH content and altered thiol-redox homeostasis in SM induced by strenuous high-intensity exercise; (2) an adaptive elevation of basal GSH level following the short-term training period; (3) an adaptive decrease of basal GSH level following 2.5 days recovery from training; (4) evidence of a relationship between the SM fibre type, physical performance capacity and GSH turnover during acute bouts of exercise; and (5) no evident effect of the level of carbohydrate intake on metabolism of GSH or energy intermediates. Furthermore, the induction of acute oxidative stress in exercising human SM and the adaptive responses to training are suggested to provide a protective antioxidant phenotype to the exercising SM during periods with repeated intense intermittent training.

The natriuretic hormone dopamine and the antinatriuretic hormone noradrenaline, acting on alpha-adrenergic receptors, have been shown to bidirectionally modulate the activity of renal tubular Na+,K+-adenosine triphosphate (ATPase). Here we have examined whether intracellular sodium concentration influences the effects of these bidirectional forces on the state of phosphorylation of Na+,K+-ATPase. Proximal tubules dissected from rat kidney were incubated with dopamine or the alpha-adrenergic agonist, oxymetazoline, and transiently permeabilized in a medium where sodium concentration ranged between 5 and 70 mM. The variations of sodium concentration in the medium had a proportional effect on intracellular sodium. Dopamine and protein kinase C (PKC) phosphorylate the catalytic subunit of rat Na+,K+-ATPase on the Ser23 residue. The level of PKC induced Na+,K+-ATPase phosphorylation was determined using an antibody that only recognizes Na+,K+-ATPase, which is not phosphorylated on its PKC site. Under basal conditions Na+,K+-ATPase was predominantly in its phosphorylated state. When intracellular sodium was increased, Na+,K+-ATPase was predominantly in its dephosphorylated state. Phosphorylation of Na+,K+-ATPase by dopamine was most pronounced when intracellular sodium was high, and dephosphorylation by oxymetazoline was most pronounced when intracellular sodium was low. The oxymetazoline effect was mimicked by the calcium ionophore A23187. An inhibitor of the calcium-dependent protein phosphatase, calcineurin, increased the state of Na+,K+-ATPase phosphorylation. The results imply that phosphorylation of renal Na+,K+-ATPase activity is modulated by the level of intracellular sodium and that this effect involves PKC and calcium signalling pathways. The findings may have implication for the regulation of salt excretion and sodium homeostasis.

SummaryTwenty patients with coronary heart disease (CHD) and elevated serum lipids were randomized into 2 groups of 10 to receive encapsulated preparations of either a concentrated ethylester form of eicosapentaenoic acid (EPA) and docosa-hexaenoic acid (DHA) or corn oil in doses of 6 g per day, given double blindly for approximately two months prior to coronary bypass surgery. Lipopolysaccharide (LPS) induced monocyte thromboplastin synthesis was studied during the preoperative period and one week following surgery. The ability of n-3 fatty acids to modify tissue factor pathway inhibitor (TFPI) and tissue plasminogen activator inhibitor (PAI-1) was also evaluated along with fibrinogen and thrombin-antithrombin III (TAT) complexes.No significant changes were noted preoperatively. Monocyte reactivity, PAI-1, fibrinogen and TAT increased significantly after surgery. These changes were not modified by preoperative loading with n-3 fatty acids.

Enzymatic properties of barley alpha-amylase 1 (AMY1) are altered as a result of amino acid substitutions at subsites -5/-6 (Cys95--Ala/Thr) and +1/+2 (Met298--Ala/Asn/Ser) as well as in the double mutants, Cys95--Ala/Met298--Ala/Asn/Ser. Cys95--Ala shows 176% activity towards insoluble Blue Starch compared to wild-type AMY1, kcat of 142 and 211% towards amylose DP17 and 2-chloro-4-nitrophenyl beta-d-maltoheptaoside (Cl-PNPG7), respectively, but fivefold to 20-fold higher Km. The Cys95--Thr-AMY1 AMY2 isozyme mimic exhibits the intermediary behaviour of Cys95--Ala and wild-type. Met298--Ala/Asn/Ser have slightly higher to slightly lower activity for starch and amylose, whereas kcat and kcat/Km for Cl-PNPG7 areor = 30% andor = 10% of wild-type, respectively. The activity of Cys95--Ala/Met298--Ala/Asn/Ser is 100-180% towards starch, and the kcat/Km is 15-30%, and 0.4-1.1% towards amylose and Cl-PNPG7, respectively, emphasizing the strong impact of the Cys95--Ala mutation on activity. The mutants therefore prefer the longer substrates and the specificity ratios of starch/Cl-PNPG7 and amylose/Cl-PNPG7 are 2.8- to 270-fold and 1.2- to 60-fold larger, respectively, than of wild-type. Bond cleavage analyses show that Cys95 and Met298 mutations weaken malto-oligosaccharide binding near subsites -5 and +2, respectively. In the crystal structure Met298 CE and SD (i.e., the side chain methyl group and sulfur atom) are near C(6) and O(6) of the rings of the inhibitor acarbose at subsites +1 and +2, respectively, and Met298 mutants prefer amylose for glycogen, which is hydrolysed with a slightly lower activity than by wild-type. Met298 AMY1 mutants and wild-type release glucose from the nonreducing end of the main-chain of 6"'-maltotriosyl-maltohexaose thus covering subsites -1 to +5, while productive binding of unbranched substrate involves subsites -3 to +3.

The interaction of the two binding sites of the starch-binding domain (SBD) of Aspergillus niger glucoamylase 1 (GA-I) with substrate has been investigated by using atomic force microscopy (AFM) and UV difference spectroscopy in combination with site-specific mutants of both SBD and GA-I. The SBD possesses two binding sites with distinct affinities towards the soluble linear substrate maltoheptaose; dissociation constants (K(d)) of 17 and 0.95 microM were obtained for W563 K (binding site 2 mutant) and W590 K (binding site 1 mutant), respectively, compared to an apparent K(d) of 23 microM for the wild-type SBD. Further, the two sites are almost but not totally independent of each other for binding, since abolishing one site does not prevent the amylose chain binding to the other site. Using AFM, we show that the amylose chains undergo a conformational change to form loops upon binding to the SBD, using either the recombinant wild-type SBD or a catalytically inactive mutant of GA-I. This characteristic conformation of amylose is lost when one of the SBD binding sites is eliminated by site-directed mutagenesis, as seen with the mutants W563 K or W590 K. Therefore, although each binding site is capable of simple binding to a ligand, both sites must be functional in order to induce a gross conformational change of the amylose molecules. Taken together these data suggest that for the complex with soluble amylose, SBD binds to a single amylose chain, site 1 being responsible for the initial recognition of the chain and site 2 being involved in tighter binding, leading to the circularisation of the amylose chain observed by AFM. Binding of the SBD to the amylose chain results in a novel two-turn helical amylose complex structure. The binding of parallel amylosic chains to the SBD may provide a basis for understanding the role of the SBD in facilitating enzymatic degradation of crystalline starches by glucoamylase 1.

The relative specificity and bond cleavage pattern of barley alpha-amylase 1 (AMY1) were dramatically changed by mutation in F(286)VD that connected beta-strand 7 of the catalytic (beta/alpha)(8)-barrel to a succeeding 3(10)-helix. This conserved tripeptide of the otherwise variable beta --alpha segment 7 lacked direct ligand contact, but the nearby residues His290 and Asp291 participated in transition-state stabilization and catalysis. On the basis of sequences of glycoside hydrolase family 13, a biased random mutagenesis protocol was designed which encoded 174 putative F(286)VD variants of C95A-AMY1, chosen as the parent enzyme to avoid inactivating glutathionylation by the yeast host. The FVG, FGG, YVD, LLD, and FLE mutants showed 12-380 and 1.8-33% catalytic efficiency (k(cat)/K(m)) toward 2-chloro-4-nitrophenyl beta-D-maltoheptaoside and amylose DP17, respectively, and 0.5-50% activity for insoluble starch compared to that of C95A-AMY1. K(m) and k(cat) were decreased 2-9- and 1.3-83-fold, respectively, for the soluble substrates. The starch:oligosaccharide and amylose:oligosaccharide specificity ratios were 13-172 and 2.4-14 for mutants and 520 and 27 for C95A-AMY1, respectively. The FVG mutant released 4-nitrophenyl alpha-D-maltotrioside (PNPG(3)) from PNPG(5), whereas C95A-AMY1 produced PNPG and PNPG(2). The mutation thus favored interaction with the substrate aglycon part, while products from PNPG(6) reflected the fact that the mutation restored binding at subsite -6 which was lost in C95A-AMY1. The outcome of this combined irrational and rational protein engineering approach was evaluated considering structural accommodation of mutant side chains. FVG and FGG, present in the most active variants, represented novel sequences. This emphasized the worth of random mutagenesis and launched flexibility as a goal for beta --alpha loop 7 engineering in family 13.

Thirty percent of all patients that consult their primary care physicians for pain in their musculoskeletal systems have pain in the neck/shoulder region, while 23% have lower back pain. Of major concern in Sweden is a patient's lengthy waiting period before seeing a specialist, with a resultant delay in treatment. A multidisciplinary group of physicians and physical therapists has therefore been working on and refining a system of tools to provide an "unbroken chain of care" for the diagnosis and treatment of patients with shoulder pathology. An unbroken chain of care enables the clinician regardless of specialty to provide the highest level of care to all patients with the same initial diagnostic and treatment parameters. Axelina is the tool that provides all the standardized treatment algorithms, rehabilitation plans and detailed home exercises. Axelina also contains a two-day instructional package with case studies presented on video.

The aim of the present study was to see whether a moderate dose of omega-3 fatty acids (FA) potentiates the beneficial effects of statins on the high risk for coronary heart disease (CHD) in patients with combined hyperlipemia.In the present double-blind parallel study, 42 patients with combined hyperlipemia with serum triglycerides 2-15 mmol/L-1 and serum total cholesterol5.3 mmol/L-1 at the end of a three-month dietary run-in period were treated with 10 mg/d atorvastatin for 10 or more weeks. During the last 5 weeks they were randomized into two groups that received either 1.68 g/d omega-3 FA as ethylesters of eicosapentaenoic (45%) and docosahexaenoic acids (39%), or placebo (corn oil). As expected, atorvastatin significantly reduced serum total LDL-cholesterol (LDL-C), triglycerides and apolipoproteins B, E, CII and CIII, whereas HDL-cholesterol (HDL-C) was increased. Addition of omega-3 FA further increased HDL-C (p0.03), and reduced systolic blood pressure (0.03), while the small dense LDL-particles (LDL III) (p0.05) and postprandial hypertriglyceridemia (p0.01) were reduced compared with the baseline, though there were no significant differences to the placebo group. This may be related to the large individual variation in these parameters and the small number of patients. No significant effects on basic or postheparin activities of lipoprotein lipase or hepatic lipase were observed after atorvastatin with or without addition of omega-3 FA.This study indicates that addition of a low dose of omega-3 FA may further improve the risk profile for CHD in patients with combined hyperlipemia treated with atorvastatin. The effect is related to reduction of postprandial hyperlipemia and redistribution of LDL subfractions.

Heparin is known to exert its antithrombotic effects by accelerating the effect of antithrombin (AT) and by mobilizing tissue factor pathway inhibitor (TFPI) into the circulation from vascular endothelium. Heparin treatment has been reported to decrease FVIIa activity by 40%; this was suggested as a new antithrombotic action of heparins. The present study was conducted to investigate whether the apparent reduction in FVIIa activity induced by unfractionated heparin (UFH) infusion in vivo is due to interactions between AT and TFPI with the FVIIa assay or due to an actual decrease in FVIIa. Blocking plasma TFPI in affinity purified anti-TFPI IgG caused a 25% increase in plasma FVIIa activity (Staclot VII - rTF, Diagnostica Stago, Aswiéres-sur-Seine, France). In vitro heparinization of plasma caused a dose-dependent decrease in FVIIa (up to 56 +/- 8%) at high heparin concentrations (1.0-5.0 IU/mL UFH), a reduction abolished by Hexadimethine Bromide (HDB) to neutralize heparin-induced activation of AT. Thus, heparin-induced activation of AT is apparently responsible for decreased FVIIa under in vitro conditions. Bolus injection followed by continuous infusion of heparin to healthy volunteers was accompanied by a prompt 50% reduction in FVIIa activity, which was sustained throughout heparin infusion and normalized within 24 hours after discontinuation of treatment. Addition of anti-TFPI IgG to postheparin plasma reversed the heparin-induced reduction in FVIIa by approximately 50%, and combined pretreatment of postheparin plasma with anti-TFPI IgG and HDB brought FVIIa to preheparin levels. The present study shows that the FVIIa assay is sensitive to TFPI and AT, especially during heparin treatment, and thereby indicates that the heparin-induced decrease in FVIIa is affected by interactions between TFPI and AT with the FVIIa assay.

Treatment with titanium dental implants is in general successful. However, an unknown number of implants do not integrate and are removed either by exfoliation or at the time of second stage surgery. It would be of importance to identify subjects at risk and predict early implant failure.In a retrospective study serum IgG antibody titers and avidity in sera from 40 subjects who had experienced titanium dental implant treatments with non-osseo-integration as the outcome (NOTI) and in sera from 40 age and gender matched control subjects who had received successful titanium dental implants (SOTI) were studied. Serum IgG titers to whole cell Actinomyces viscosus, Bacteroides forsythus, Porphyromonas gingivalis, Staphylococcus aureus, and Streptococcus intermedius sonicated antigen preparations were studied by ELISA.Serum IgG antibody titers to S. aureus were significantly higher in subjects with SOTI than in NOTI (p0.001) suggesting that higher titers indicate protection against implant failure as a result of S. aureus infection. Statistically significant higher serum IgG antibody avidity to P. gingivalis and B. forsythus were found in subjects with SOTI than in subjects with NOTI (p0.01 and p0.001, respectively). Statistical analysis failed to demonstrate antibody titer or avidity differences to the other pathogens studied. The likelihood that SOTI was associated with a high OD reading for S. aureus was 13.1:1 (p0.001). Whether subjects were edentulous or not, or if they had lost teeth because of periodontitis or caries did not seem to matter.Serum IgG antibodies relative to B. forsythus, P. gingivalis and S. aureus may be associated with the outcome of implant procedures and explain why early implant failures occur.

Anaemia in haemodialysis patients can be effectively treated with erythropoietin. We investigated whether subcutaneous (SC) epoetin ss administered once weekly was as effective as the same weekly dosage given in two to three divided doses.One hundred and fifty-eight patients (delivered Kt/V1.0, where K=dialyser-renal urea clearance, t=dialysis time and V=filtration volume, obtained by urea kinetic modelling) were randomized to treatment with SC epoetin beta either once weekly (n=118), or to their original dosage two or three times weekly (control group, n=40) for 24 weeks. All patients received intravenous iron supplementation when necessary.Eight-eight patients in the once weekly group and 30 patients in the control group were treated for at least 16 weeks and are included in the analysis. Stable haemoglobin levels were maintained without epoetin dose increases in 73% of patients in both groups. Mean haemoglobin levels at randomization and after 16 and 24 weeks were 11.4, 11.1 and 11.1 g/dl, respectively, in the once weekly group compared with 11.2, 11.3 and 11.2 g/dl, respectively, in the control group. The mean weekly epoetin beta dosages at randomization and after 16 and 24 weeks were 102, 103 and 106 IU/kg bodyweight, respectively, in the once weekly group compared with 109, 109 and 115 IU/kg bodyweight, respectively, in the control group. No statistically significant between-group differences were apparent for changes in haemoglobin levels or epoetin beta dosages at week 24.Once weekly SC administration of epoetin beta is as safe and effective in maintaining haemoglobin levels in stable haemodialysis patients as two or three times weekly administration of the same total dose. By using the once weekly regimen, patients can avoid up to 104 injections per year. This would reduce clinic time for patients who do not self administer, and may also encourage self-administration and improve overall compliance.

In a prospective study, 105 children with juvenile chronic arthritis (JCA) were clinically and radiographically examined. The aim was to investigate the distribution of symptoms and clinical signs of temporomandibular disorders and to study correlations with radiographic mandibular condylar lesions. The present material appeared to be a representative sample of Swedish JCA children with respect to the distribution of genders and JCA subtypes as well as the peak of onset of the disease. Symptoms from the masticatory system were common (26%) and pain at mandibular function and stiffness at mouth opening the most frequent ones. Restricted maximal voluntary mouth opening (MVM) was the most frequent clinical finding. Radiographic condylar lesions, frequently found in the present material (39%), were significantly correlated with postnormal occlusion, restricted MVM, anterior open bite and mandibular retrognathia. However, most of the JCA children with radiographic condylar lesions did not present with symptoms from the masticatory system, postnormal occlusion, restricted MVM, anterior open bite or mandibular retrognathia.

To assess criteria for individual response and remission based on the Disease Activity Score (DAS) in patients with RA participating in a long-term observational study.The EULAR (European League against Rheumatism) criteria for individual response and a recently proposed remission criterion, DAS1.6, were applied to 90 patients with RA after treatment for 2 yr with disease-modifying anti-rheumatic drugs (DMARDs) and/or corticosteroids.Seventy-six per cent of the patients were classified as responders (46% good and 30% moderate responders). Good responders had significantly more improvement in pain and Health Assessment Questionnaire (HAQ) score than moderate responders and non-responders. Paired comparisons showed significant X-ray progression both for moderate responders and non-responders but not for good responders. Twenty-nine per cent of all responders had an end-point DAS2.4, indicating active disease. In this group of responders, X-ray changes progressed significantly, but this could not be demonstrated in the group of responders with DASor = 2.4. Thirty-six per cent of the patients included in the study were classified as being in remission after 2 yr of treatment. The group of patients in remission showed no evidence of X-ray progression after 2 yr.Response and remission criteria based on DAS were useful in a study of patients with RA who were managed essentially as in clinical practice. The criteria used showed construct and criterion validity, although discriminant validity could not be shown. The application of valid criteria for response and remission in clinical practice may be a useful aid in the evaluation of treatment effects and in making treatment decisions for individual patients.

TFPI is a potent inhibitor of the extrinsic coagulation system constitutively synthesized by endothelial cells. A major portion of intravascular TFPI is stored associated with endothelial cells. and administration of unfractionated heparin (UFH) in vivo causes a prompt mobilization of TFPI into the circulation. The present study was conducted to investigate how UFH affected the synthesis, secretion and anticoagulant potency of TFPI in endothelial cells in vitro. A spontaneously transformed immortal endothelial cell line was used (ECV304). Stimulation of ECV304 cells with UFH caused a prompt dose-dependent (0-5 IU UFH/ml) release of TFPI to the medium accompanied by no change of TFPI at the surface membrane assessed by immunocytochemical methods. Northern blot analysis revealed two mRNA transcripts for TFPI with a molecular size of 1.4 kb and 4.4 kb, respectively. Stimulation of ECV304 cells for 24 hrs with various concentrations of UFH caused a dose-dependent increase of TFPI in the medium (6.2-29.6 ng/10(6) cells within the concentration range 0-10 IU/ml). A similar dose-dependent increase in the expression of both TFPI mRNA species was observed. Long-term incubation of ECV304 cells with 5.0 IU/ml UFH caused a 5-10 fold increase in the TFPI concentration accumulated in the medium over 48 hrs. The increased TFPI mRNA expression induced by UFH appeared already after 10 min, peaked after 2-4 hrs, remained augmented throughout the entire period of UFH exposure, and preceded the synthesis-dependent increase in TFPI release by 2-4 hrs. The procoagulant activity of the cells was downregulated by 36% and the contribution of TFPI to the anticoagulant potency of ECV304 cells was moderately increased after 24 hrs heparin stimulation. It is suggested that these mechanisms are of major importance for the anticoagulant function of heparins.

Several laser procedures, extracorporeal lithotripsies (ESWL), and high-velocity missile trauma generate pressure transients that are transmitted through the tissues. Despite several publications demonstrating shock wave-induced tissue injury, little is known about its pathophysiology. This study introduces an in vitro model for studying shock wave effects on endothelial cell (EC) monolayers.A Nd:YAG laser-driven flyer-plate technique was used to generate shock waves. Physical characteristics were determined with a pressure transducer, a high-speed video camera, and sequential photography. Biological effects were studied with phase contrast and lightfield microscopy, computerized morphometry, immunocytochemistry, spectrophotometry, and enzyme-linked immunosorbent assay (ELISA).The shock waves generated were highly reproducible. Cavitation was verified and quantified, and its extent could be varied in the vials. Exposed cultures exhibited areas with cell membrane damage and cell detachment. Release of LD was elevated (P0.01) in exposed vials. The EC lesions were larger (P0.01) in cultures submitted to high vs. low extent of cavitation.The flyer-plate model can be used to subject cell monolayers to defined and reproducible shock waves causing immediate cell injury similar to the previously reported vascular lesions associated with ESWL, pulsed lasers, and blast trauma. With the flyer-plate model, such lesions may be further studied on the cellular and subcellular levels.

The transformation of psychiatry during the last decades has also resulted in changed and more independent roles for mental health workers of all categories. The nurse has traditionally had a central role in psychiatric care. In the community oriented care the nurse still plays an important role which can be summarised in four points: 1. focus on the rehabilitation of long term mentally ill patients, 2. the case manager function, 3. to provide individual support in the daily living and 4. to provide support to the relatives of the patients. Those different roles are described. The research being done on these different approaches is presented and it is concluded that case management, social skills training and family intervention programmes have all proved to be effective. In these programmes nurses often play a central role.

Svensson Tony B. De l'intérêt d'un carnet d'entraînement. In: Les Cahiers de l'INSEP, n°24, 1999. 2e Congrès international de triathlon de l’INSEP – 2nd INSEP international triathlon Congress pp. 146-147.

We have used adult mouse superior cervical ganglia (SCG) to study the role of mitogen activated protein (MAP) kinase activity during axonal outgrowth in vitro. An initial peak in activity within the first hours of culture was followed by a substantially higher activity after 1 to 2 days, a time when axons were actively growing. The latter peak is probably a result of both higher levels of protein and increased activity. The addition of nerve growth factor stimulated both outgrowth and kinase activity, whereas treating the cultures with the kinase inhibitor PD98059 had an opposite effect. Taken together, the results suggest that activation of the MAP kinase pathway could be involved in the initiation as well as regulation of axonal outgrowth from adult SCG.

The authors determined the levels of the secreted, interstitially located extracellular superoxide dismutase (EC-SOD), the cytosolic copper- and zinc-containing SOD (CuZn-SOD), and the mitochondrial manganese-containing SOD (Mn-SOD) in tears, cornea, sclera, aqueous humor, lens, vitreous body, and retina of the human eye.SOD enzymatic activity was determined by the direct spectrophotometric method using KO2. The amounts of EC-SOD and CuZn-SOD protein were also determined using enzyme-linked immunosorbent assay.Tears, aqueous humor, and vitreous body contained little SOD activity. Here ascorbate is the major scavenger of superoxide radicals. Of the ocular tissues studied, the lens contained the least SOD activity, 15-fold less than that of the average human tissue. The majority came from CuZn-SOD, but the lens also contained considerable amounts of inactive enzyme. There was some Mn-SOD, but the EC-SOD content was negligible. The cornea and sclera contained several times more SOD activity than the lens, and the EC-SOD contents were equal to or larger than those of CuZn-SOD. The retina contained much CuZn-SOD activity, but the EC-SOD and Mn-SOD activities were low.Ocular tissues and fluids, except for those in the retina, are less protected against superoxide radicals than extraocular tissues and fluids. The large amounts of EC-SOD in the sclera and cornea may be related to the risk for photochemical production of superoxide in these tissues. If inflammatory reactions occur, ocular tissues are at risk for damage induced by superoxide radicals and peroxynitrite, the reaction product with nitric oxide.

The onset of action rate of CPH-82 (Reumacon®), was compared with that of auranofin (AUR; Ridaura®) in a 36-week randomised, double-blind, multicentre study of 60 patients with rheumatoid arthritis (RA). As compared with respective baseline values, the CPH-82 group manifested significant improvement in grip strength, Ritchie's articular index (RAI), pain ratings, and HAQ (health assessment questionnaire) scores after 8, 12, 24, and 36 weeks of treatment, with the exception of the 24-week HAQ score. The AUR group manifested corresponding improvement in RAI after 8, 12, 24, and 36 weeks. Significant differences in changes from baseline values in favour of the CPH-82 group were found for grip strength at 12 and 24 weeks, RAI score at 8 weeks, VAS score at 8 and 12 weeks, and HAQ score at 8 weeks. The findings suggest CPH-82 to manifest a more rapid onset of action than AUR. The two drugs were similar in tolerance and safety. O. Bjorneboe, Dept. of Rheumatology, Martina Hansens Hospital, P. O. Box 23, N-1355 ...

SWEDEC is now organised as described in this paper and the main tasks are to: support Swedish international commitments, both for military and humanitarian purposes, in the field of EOD and demining, by providing training for involved units and individuals as well as developing methods and equipment; train regular and reserve officers, police officers and civilian personnel; and develop the function of military engineering.

Differences in sex ratios have since long been recognized in several common chronic diseases and are particularly common in rheumatic diseases, inflammatory as well as non-inflammatory. The present presentation will focus on gender in some of the most frequent rheumatic disorders - rheumatoid arthritis, systemic lupus erythematosus and ankylosing spondylitis. The predictive ability of gender in these common rheumatic diseases will be reviewed and possible confounding issues considered. Rheumatoid arthritis (RA) In RA the female:male ratio is about 2-3:1. Already in the year 1800 Augustin Jacob Landre-Beauvais described the female preponderance in a group of patients who probably had RA. Since long women are believed to run a more serious disease course than men. In recent years, several long-term studies have shown that women have higher DAS28, more pain, worse HAQ and lower remission rates compared with men. However, observational studies over 2, 5 and 8 years have shown that joint damage assessed by radiographic scoring was similar in men and women. This inconsistent situation may be explained by the fact that pain, unrelated to the inflammatory process, confounds the interpretation of the values obtained for DAS or DAS28. This may give rise to overestimation of the disease activity and possibly result in suboptimal treatment. A common cause of such non-inflammatory pain is fibromyalgia (FM), a common co-morbidity in RA with pronounced female preponderance (about 9:1). FM in patients with RA has been found to be an independent predictor of DAS28 and may therefore confound the evaluation of disease activity. Systemic lupus erythematosus (SLE) SLE displays a striking female predominance, 9:1 in childbearing age. Of interest, men seem to be at risk of a worse outcome compared with women. Thus, male sex has turned out to be a predictor of serious kidney involvement. Moreover, disease severity including more cardiovascular and renal disease was reported to be more pronounced in men. Furthermore, in childhood SLE, boys were reported to have a higher prevalence of severe renal disease and poorer outcome. These studies suggest that gender is a predictor of both disease expression and severity and should be taken into account in the often difficult choice of treatment strategy in patients with SLE. Ankylosing spondylitis (AS) AS has a male preponderance estimated to about 3:1. Gender differences in disease profile and disease course have been described with more low back spondylitis and more radiographic changes in men and more involvement of neck and peripheral joints in women. Furthermore, studies have suggested that women have higher disease activity and develop more functional impairment than men. However, further studies using different assessment methods produced diverging results. Moreover, active disease, measured by objective markers, and male gender predicted benefit from biologic drugs while active disease, assessed by subjective markers, and female gender did not. Thus, gender may predict disease expression and response to therapy in AS although the evaluation of disease activity may be confounded by the available assessment methods. Conclusions In rheumatic diseases gender differences in disease prevalence are striking. Gender may predict disease expression, course and outcome and should be taken into account in clinical trials and observational studies as well as in clinical practice. In addition, the possibility that assessment instruments containing non-disease specific components may confound the evaluation of the disease should be acknowledged and stimulate to the development of more specific instruments. Disclosure of Interest None Declared