Dietary omega-3 fatty acids modulate the production of platelet-derived microvesicles in an in vivo inflammatory arthritis model.

PURPOSE: The aim of this study was to investigate the effects of different ω-3 polyunsaturated fatty acid (PUFA) enriched diets, including a novel renewable plant source of ω-3 fatty acids (Buglossoides arvensis), on the development and progression of rheumatoid arthritis (RA). METHODS: RA was induced in mice consuming experimental diets using the K/BxN model. The experimental diets consisted of either a western control diet (control), diets containing B. arvensis oil or fish oil. The effects of the diets on platelets, platelet microvesicles (PMVs), and inflammatory markers such as clinical index, ankle thickness and cytokine/chemokine release were measured. RESULTS: While ω-3 PUFA-enriched diets did not prevent the development of arthritis in the K/BxN model, a significant decrease in ankle swelling was observed compared to the control group. Platelets isolated from mice consuming either low content of B. arvensis oil or fish oil diets exhibited significantly decreased PMVs production compared to mice consuming the control diet. CONCLUSION: Our study provides insight into the contribution of ω-3 PUFA supplementation in modulating the pro-inflammatory phenotype of platelets in RA pathology. Furthermore, our study suggests that low concentrations of dietary B. arvensis oil may have similar anti-inflammatory potential seen with dietary fish oil supplementation.

Single and multiple inhibitors of the biosynthesis of 5-, 12-, 15-lipoxygenase products derived from cinnamyl-3,4-dihydroxy-α-cyanocinnamate: Synthesis and structure-activity relationship.

The involvement of lipoxygenases in various pathologies, combined with the unavailability of safe and effective inhibitors of the biosynthesis of their products, is a source of inspiration for the development of new inhibitors. Based on a structural analysis of known inhibitors of lipoxygenase products biosynthesis, a comprehensive structure-activity study was carried out, which led to the discovery of several novel compounds (16a-c, 17a) demonstrating promising potency to inhibit the biosynthesis of products of 5-, 12- and 15-LO. Compounds 16b and 16c outperformed zileuton (1), the only FDA-approved 5-LO inhibitor, as well as known inhibitors such as caffeic acid phenethyl ester (CAPE (2)) and cinnamyl-3,4-dihydroxy-α-cyanocinnamate (CDC (4)). However, the introduction of a cyano group at the α-position of the carbonyl abolished the activity. Compounds 16a and 17a also inhibited the biosynthesis of 12- and 15-LO products. Compounds 16a, 17a far surpassed baicalein, a known 12-LO inhibitor, as inhibitors of 12-LO products biosynthesis. Compound 17a and CDC (4) showed equivalent inhibition of LO products, proposing that the double bond in the ester moiety is not necessary for the inhibitory activity. The introduction of the cyano group, as in compound 17a, at the α-position of the carbonyl in compound 16a significantly reduced the inhibitory activity against the biosynthesis of 15-LO products. In addition to the interactions with residues His372 and Phe421 also found with zileuton and CAPE, compounds 16a and 16c each interact with residue His367 as shown by molecular docking. This new interaction may explain their high affinity with the 5-LO active site.

Algerian Propolis from Distinct Geographical Locations: Chemical Profiles, Antioxidant Capacity, Cytotoxicity and Inhibition of 5-Lipoxygenase Product Biosynthesis.

Propolis was collected from honeybee hives in three geographically distinct Algerian climates and extracts were characterized for composition and bioactivity. Bees were identified as native subspecies using an in-silico DraI mtDNA COI-COII test. Over 20 compounds were identified in extracts by LC-MS. Extracts from the Medea region were more enriched in phenolic content (302±28 mg GAE/g of dry extract) than those from Annaba and Ghardaia regions. Annaba extracts had the highest flavonoid content (1870±385 mg QCE/g of dry extract). Medea extracts presented the highest free-radical scavenging activity (IC50=13.5 µg/mL) using the DPPH radical assay while Ghardaia extracts from the desert region were weak (IC50>100 µg/mL). Antioxidant activities measured using AAPH oxidation of linoleic acid were similar in all extracts with IC50 values ranging from 2.9 to 4.9 µg/mL. All extracts were cytotoxic (MTT assay) and proapoptotic (Annexin-V) against human leukemia cell lines in the low µg/mL range, although the Annaba extract was less active against the Reh cell line. Extracts inhibited cellular 5-lipoxygenase product biosynthesis with IC50 values ranging from 0.6 to 3.2 µg/mL. Overall, examined propolis extracts exhibited significant biological activity that warrant further characterization in cellular and in vivo models.

EGCG inhibits the inflammation and senescence inducing properties of MDA-MB-231 triple-negative breast cancer (TNBC) cells-derived extracellular vesicles in human adipose-derived mesenchymal stem cells.

BACKGROUND: Triple-negative breast cancer (TNBC) cells' secretome can induce a pro-inflammatory phenotype in human adipose-derived mesenchymal stem cells (hADMSC). This can be prevented by the green tea polyphenol epigallocatechin-3-gallate (EGCG). The impact of EGCG on the paracrine regulation that the extracellular vesicles (EVs) specifically exert within the TNBC secretome remains unknown. METHODS: EVs were obtained from a TNBC-derived serum-starved MDA-MB-231 cell model treated or not with EGCG under normoxic or hypoxic (< 1% O2) culture conditions. RNA-Seq analysis was used to assess the EVs' genetic content. The modulation of inflammatory and senescence markers in hADMSC was evaluated by RT-qPCR using cDNA arrays and validated by immunoblotting. A protein profiler phospho-kinase array was used to explore signaling pathways. RESULTS: While hypoxic culture conditions did not significantly alter the genetic content of MDA-MB-231-secreted EVs, the addition of EGCG significantly modified EVs genetic material at low oxygen tension. Gene expression of cancer-associated adipocyte pro-inflammatory markers CXCL8, CCL2 and IL-1ß was increased in hADMSC treated with EVs. Concomitantly, EVs isolated from MDA-MB-231 treated with EGCG (EGCG-EVs) downregulated CCL2 and IL-1ß, while inducing higher expression of CXCL8 and IL-6 levels. EVs activated CHK-2, c-Jun, AKT and GSK-3ß signaling pathways in hADMSC, whereas EGCG-EVs specifically reduced the latter two as well as the serum starvation-induced senescence markers p21 and ß-galactosidase. Finally, the mitochondrial content within the TNBC cells-derived EVs was found reduced upon EGCG treatment. CONCLUSION: This proof of concept study demonstrates that the chemopreventive properties of diet-derived polyphenols may efficiently target the paracrine regulation that TNBC cells could exert upon their surrounding adipose tissue microenvironment.

Caffeic acid phenethyl ester analogues as selective inhibitors of 12-lipoxygenase product biosynthesis in human platelets.

The inflammatory response is an essential process for the host defence against pathogens. Lipid mediators are important in coordinating the pro-inflammatory and pro-resolution phases of the inflammatory process. However, unregulated production of these mediators has been associated with chronic inflammatory diseases such as arthritis, asthma, cardiovascular diseases, and several types of cancer. Therefore, it is not surprising that enzymes implicated in the production of these lipid mediators have been targeted for potential therapeutic approaches. Amongst these inflammatory molecules, the 12-hydroxyeicosatetraenoic acid (12(S)-HETE) is abundantly produced in several diseases and is primarily biosynthesized via the platelet's 12-lipoxygenase (12-LO) pathway. To this day, very few compounds selectively inhibit the 12-LO pathway, and most importantly, none are currently used in the clinical settings. In this study, we investigated a series of polyphenol analogues of natural polyphenols that inhibit the 12-LO pathway in human platelets without affecting other normal functions of the cell. Using an ex vivo approach, we found one compound that selectively inhibited the 12-LO pathway, with IC50 values as low as 0.11 µM, with minimal inhibition of other lipoxygenase or cyclooxygenase pathways. More importantly, our data show that none of the compounds tested induced significant off-target effects on either the platelet's activation or its viability. In the continuous search for specific and better inhibitors targeting the regulation of inflammation, we characterized two novel inhibitors of the 12-LO pathway that could be promising for subsequent in vivo studies.

Molecular characterization of the anticancer properties associated with bee venom and its components in glioblastoma multiforme.

Glioblastoma multiforme (GBM) is a frequent form of malignant glioma. Strategic therapeutic approaches to treat this type of brain tumor currently involves a combination of surgery, radiotherapy and chemotherapy. Nevertheless, survival of GBM patients remains in the 12-15 months range following diagnosis. Development of novel therapeutic approaches for this malignancy is therefore of utmost importance. Interestingly, bee venom and its components have shown promising anti-cancer activities in various types of cancer even though information pertaining to GBMs have been limited. The current work was thus undertaken to better characterize the anti-cancer properties of bee venom and its components in Hs683, T98G and U373 human glioma cells. MTT-based cell viability assays revealed IC50 values of 7.12, 15.35 and 7.60 µg/mL for cell lines Hs683, T98G and U373 treated with bee venom, respectively. Furthermore, melittin treatment of these cell lines resulted in IC50 values of 7.77, 31.53 and 12.34 µg/mL, respectively. Cell viability assessment by flow cytometry analysis confirmed signs of late apoptosis and necrosis after only 1 h of treatment with either bee venom or melittin in all three cell lines. Immunoblotting-based quantification of apoptotic markers demonstrated increased expression of Bak and Bax, while Caspsase-3 levels were significantly lower when compared to control cells. Quantification by qRT-PCR showed increased expression levels of long non-coding RNAs RP11-838N2.4 and XIST in glioma cells treated with either bee venom or melittin. Overall, this study provides preliminary insight on molecular mechanisms via which bee venom and its main components can impact viability of glioma cells and warrants further investigation of its anticancer potential in gliomas.

Evaluating the mitochondrial activity and inflammatory state of dimethyl sulfoxide differentiated PLB-985 cells.

Neutrophils play a key role in the innate immunity with their ability to generate and release inflammatory mediators that promote the inflammatory response and consequently restore the hemostasis. As active participants in several steps of the normal inflammatory response, neutrophils are also involved in chronic inflammatory diseases such as asthma, atherosclerosis, and arthritis. Given their dual role in the modulation of inflammation, regulating the inflammatory response of neutrophils has been suggested as an important therapeutic approach by numerous researchers. The neutrophils have a relatively short lifespan, which can be problematic for some in vitro experiments. To address this issue, researchers have used the human monomyelocyte cell line PLB-985 as an in vitro model for exploratory experiments addressing neutrophil-related physiological functions. PLB-985 cells can be differentiated into a neutrophil-like phenotype upon exposure to several agonists, including dimethyl sulfoxide (DMSO). Whether this differentiation of PLB-985 affects important features related to the neutrophil's normal functions (i.e., mitochondrial activity, eicosanoid production) remains elusive, and characterizing these changes will be the focal point of this study. Our results indicate that the differentiation affected the proliferation of PLB-985 cells, without inducing apoptosis. A significant decrease in mitochondrial respiration was observed in differentiated PLB-985 cells. However, the overall mitochondria content was not affected. Immunoblotting with mitochondrial antibodies revealed a strong modulation of the succinate dehydrogenase A, superoxide dismutase 2, ubiquinol-cytochrome c reductase core protein 2 and ATP synthase subunit α in differentiated PLB-985 cells. Finally, eicosanoids (leukotriene B4, 12-hydroxyheptadecatrienoic and 15-hydroxyeicosatetraenoic acids) production was significantly increased in differentiated cells. In summary, our data demonstrate that the differentiation process of PLB-985 cells does not impact their viability despite a reduced respiratory state of the cells. This process is also accompanied by modulation of the inflammatory state of the cell. Of importance, our data suggest that PLB-985 cells could be suitable in vitro candidates to study mitochondrial-related dysfunctions in inflammatory diseases.

New Zileuton-Hydroxycinnamic Acid Hybrids: Synthesis and Structure-Activity Relationship towards 5-Lipoxygenase Inhibition.

A novel series of zileuton-hydroxycinnamic acid hybrids were synthesized and screened as 5-lipoxygenase (5-LO) inhibitors in stimulated HEK293 cells and polymorphonuclear leukocytes (PMNL). Zileuton's (1) benzo[b]thiophene and hydroxyurea subunits combined with hydroxycinnamic acid esters' ester linkage and phenolic acid moieties were investigated. Compound 28, bearing zileuton's (1) benzo[b]thiophene and sinapic acid phenethyl ester's (2) α,ß-unsaturated phenolic acid moiety 28, was shown to be equipotent to zileuton (1), the only clinically approved 5-LO inhibitor, in stimulated HEK293 cells. Compound 28 was three times as active as zileuton (1) for the inhibition of 5-LO in PMNL. Compound 37, bearing the same sinapic acid (3,5-dimethoxy-4-hydroxy substitution) moiety as 28, combined with zileuton's (1) hydroxyurea subunit was inactive. This result shows that the zileuton's (1) benzo[b]thiophene moiety is essential for the inhibition of 5-LO product biosynthesis with our hydrids. Unlike zileuton (1), Compound 28 formed two π-π interactions with Phe177 and Phe421 as predicted when docked into 5-LO. Compound 28 was the only docked ligand that showed a π-π interaction with Phe177 which may play a part in product specificity as reported.

Extraction, Antioxidant Capacity, 5-Lipoxygenase Inhibition, and Phytochemical Composition of Propolis from Eastern Canada.

Soxhlet (SE), microwave-assisted (MAE) and ultrasound-assisted (UAE) extraction were compared using ten extraction solvents for their efficiency to extract phenolic and flavonoid antioxidants from Eastern Canada propolis. Extracts were compared for total phenolic (TPC) and total flavonoid (TFC) content, and radical scavenging activities. Anti-inflammatory activity through inhibition of 5-lipoxygenase (5-LO) products biosynthesis in HEK293 cells was also evaluated. The results showed that SE extracts using polar solvents had the highest TPC and TFC. Extracts obtained with ethanol, methanol and acetone were effective free radical scavengers, and showed 5-LO inhibition similar to zileuton. UAE was an effective extraction method since the extracts obtained were comparable to those using SE and the MAE while being done at room temperature. With UAE, extracts of less polar solvents showed similar free radical scavenging and 5-LO inhibition to extracts of much more polar solvents such as methanol or ethanol. Reversed-phase liquid chromatography tandem mass spectrometry confirmed the presence of 21 natural compounds in the propolis extracts based on the comparison of intact mass, chromatographic retention time and fragmentation patterns derived from commercial analytical standards. The current study is the first of its kind to concurrently investigate solvent polarity as well as extraction techniques of propolis.

Complex economic activities concentrate in large cities.

Human activities, such as research, innovation and industry, concentrate disproportionately in large cities. The ten most innovative cities in the United States account for 23% of the national population, but for 48% of its patents and 33% of its gross domestic product. But why has human activity become increasingly concentrated? Here we use data on scientific papers, patents, employment and gross domestic product, for 353 metropolitan areas in the United States, to show that the spatial concentration of productive activities increases with their complexity. Complex economic activities, such as biotechnology, neurobiology and semiconductors, concentrate disproportionately in a few large cities compared to less--complex activities, such as apparel or paper manufacturing. We use multiple proxies to measure the complexity of activities, finding that complexity explains from 40% to 80% of the variance in urban concentration of occupations, industries, scientific fields and technologies. Using historical patent data, we show that the spatial concentration of cutting-edge technologies has increased since 1850, suggesting a reinforcing cycle between the increase in the complexity of activities and urbanization. These findings suggest that the growth of spatial inequality may be connected to the increasing complexity of the economy.

A computed tomography study of the fibula: morphology, morphometry, intramedullary anatomy, application prospects on intramedullary nailing.

PURPOSE: The intramedullary percutaneous pinning in fractures of the lateral malleolus is a technique of osteosynthesis that can reduce complications of ORIF. Our study describes the morphology and the morphometry of the fibula, in particular intramedullary, so as to specify the best fibular nail features. METHODS: We conducted a retrospective study on CT acquisitions of fibulae in vivo. We studied total length, and the distal malleolar angle. Regarding intramedullary morphology, six axial study levels were defined. Each level was assigned a morphometric classification (oval, triangular, quadrangular or irregular), and a measure of the diameter of the cavity. The distance between the smaller diameter and the malleolar tip was investigated. RESULTS: We included 50 patients for 97 fibulae. The average age was 66.5 years. The irregular morphology type was the most frequently found. The average length was 370.5 mm (SD = 18.1; CI 95% [366.9; 374.1]), the average distal malleolar angle was 163.5° (SD = 3.7; CI 95% [162.7; 164.2]). The average minimal intramedullary diameter at malleolus level was 3.2 mm (SD = 1.2; CI 95% [3.0; 3.5]), with a minimum size reaching 95.8 mm (SD = 13.8; CI 95% [93.0; 98.5]) of the malleolar tip. CONCLUSIONS: The analysis of morphological parameters of the fibula, in particular the lateral malleolus and intramedullary morphology is necessary for the design of a morpho-adapted nail. Interpersonal variability must be taken into account by the implant industry to offer nails of suited lengths and diameters.

Pyoderma gangrenosum or necrotising fasciitis? Diagnostic and therapeutic wanderings.

A case of post-traumatic lower-limb pyoderma gangrenosum (PG) in a 77-year-old female is reported. The diagnosis of PG is frequently one of exclusion, and it is therefore unsurprising that the condition was initially mistaken for necrotising fasciitis then for necrotising bacterial dermo-hypodermitis. Medical and surgical treatment for those conditions proved ineffective. This fact, together with the atypical presentation, promoted a re-evaluation of the diagnosis. The clinical findings and investigation results converged to suggest PG, and a therapeutic trial was initiated. The good treatment response and negative findings from tests for other conditions established the diagnosis of post-traumatic PG.

Impact of muscle activation on ranges of motion during active elbow movement in children with spastic hemiplegic cerebral palsy.

BACKGROUND: Children with spastic hemiplegic cerebral palsy are restricted in their daily activities due to limited active ranges of motion of their involved upper limb, specifically at the elbow. Their impaired muscles are frequently targeted by anti-spastic treatments that reduce muscle tone. But these treatments do not necessarily improve the limb function. There is a lack of comprehensive knowledge of the quantitative relations between muscle activation and joint active ranges of motion. Consequently, the objective of this study is to quantify the impact of muscle activation on the elbow active ranges of motion. METHODS: During voluntary elbow pronation/supination and extension/flexion movements, kinematic and electromyographic measurements were collected from the involved upper limb of 15 children with spastic hemiplegic cerebral palsy (mean age=8.7 years, standard deviation=2.2) and the dominant upper limb of 15 age-matched children who are typically developing. Representative indicators of the muscle activation, such as the muscle co-activation, were extracted from the electromyographic measurements. FINDINGS: Muscle co-activation in the involved upper limb accounted for 78% and 59% of the explained variance of the supination and extension limited active ranges of motion respectively. The agonist and antagonist muscle activations were both longer in the involved upper limb. INTERPRETATIONS: This study succeeded in quantifying the impact of longer antagonist muscle activation on decreased elbow active ranges of motion in children with spastic hemiplegic cerebral palsy. Longer agonist muscle activation suggests that strengthening agonist muscles could increase the extension and supination ranges of motion, which constitutes a perspective of future clinical studies.

Comparison of four different display designs of a novel anaesthetic monitoring system, the 'integrated monitor of anaesthesia (IMA)'.

BACKGROUND: A novel monitoring system (integrated monitor of anaesthesia, IMA) which integrates three components of general anaesthesia on one single display was developed. The focus of this study was to evaluate the performance and user-friendliness of four different display designs. METHODS: Four interface displays of the IMA were developed, including one numerical, one numerical and graphical (mixed numerical-graphical), one only graphical, and one an advanced two-dimensional graphical display. Each of the four displays was evaluated in a random order by 10 staff anaesthetists and 10 residents/fellows using a set of five scenarios. Scenarios involved one or more abnormal variables that participants had to verbally phrase. For each interface test, reaction time, response accuracy, and NASA-Task Load Index were measured and compared. RESULTS: The numerical, graphical, and advanced-graphical interfaces yielded similar median reaction times, respectively, 7.99 s (5.15-10.79), 8.21 s (6.20-11.88), and 9.43 s (6.19-13.3). Reaction times were significantly shorter (P<0.006) with the mixed numerical-graphical interface: 6.26 s (4.52-8.32). The correct response rate was significantly lower in the graphical interface. The three others presented no statistical difference when compared among each other. The mixed numerical-graphical interface yielded a significantly lower NASA-TLX than the numerical and the advanced-graphical interfaces (19/100 vs 34/100, P<0.003). CONCLUSIONS: A mixed numerical-graphical display design appears to present the best results in terms of user reaction times, response accuracy, and performance index when detecting abnormal critical events.

Automatic decomposition of multichannel intramuscular EMG signals.

We describe an automatic algorithm for decomposing multichannel EMG signals into their component motor unit action potential (MUAP) trains, including signals from widely separated recording sites in which MUAPs exhibit appreciable interchannel offset and jitter. The algorithm has two phases. In the clustering phase, the distinct, recurring MUAPs in each channel are identified, the ones that correspond to the same motor units are determined by their temporal relationships, and multichannel templates are computed. In the identification stage, the MUAP discharges in the signal are identified using matched filtering and superimposition resolution techniques. The algorithm looks for the MUAPs with the largest single channel components first, using matches in one channel to guide the search in other channels, and using information from the other channels to confirm or refute each identification. For validation, the algorithm was used to decompose 10 real 6-to-8-channel EMG signals containing activity from up to 25 motor units. Comparison with expert manual decomposition showed that the algorithm identified more than 75% of the total 176 MUAP trains with an accuracy greater than 95%. The algorithm is fast, robust, and shows promise to be accurate enough to be a useful tool for decomposing multichannel signals. It is freely available at http://emglab.stanford.edu.

Properties of cryptic epitopes and their corresponding antibodies as indicated by the study of human and ovine growth hormones.

Antibodies (Ab) directed to hidden antigenic determinants (cryptotopes) are undesirable because they are not neutralizing. Additionally, we have previously demonstrated a close association between the extent of Ab to cryptic determinants and the expression of autoantibodies (autoAb) under some experimental conditions. Thus, the first objective of this work was to establish the physicochemical characteristics of Ab to cryptotopes and the second one was to examine the structural features of cryptic epitopes themselves. Using human and ovine growth hormones (hGH and oGH) as antigenic models and competition ELISA under different conditions of temperature, pH or ionic strength, we did not find any difference between the binding properties of anti-cryptic epitope antibodies (Ab) and anti-native epitope Ab. Then, using synthetic peptides and tryptic digests and direct and competition ELISAs we studied the structures of cryptic hGH and oGH epitopes. Isolated peptides either in solution or adsorbed on microplates failed to react. Partially digested hGH was recognized only when insolubilized on microplates, and anti-oGH Ab only reacted with a large fragment of the hormone either in solution or insolubilized. These results indicate that, at least in the case of hGH and oGH, cryptic epitopes are not simple linear sequences, as commonly referred without any evidence, but new exposed conformational structures different from those found in the native antigen.

Simulation of progressive spinal deformities in Duchenne muscular dystrophy using a biomechanical model integrating muscles and vertebral growth modulation.

BACKGROUND: Ninety percent of Duchenne muscular dystrophy patients develop scoliosis in parallel with evident muscular and structural impairment. Altered muscular spinal loads acting on growing vertebrae are likely to promote a self-sustaining spinal deformation process. The purpose of this study was to simulate the effect of asymmetrical fat infiltration of the erector spinae muscles combined with vertebral growth modulation over a period of growth spurt. METHODS: A finite element model of the trunk was built. It integrates (1) longitudinal growth of vertebral bodies and its modulation due to mechanical stresses, (2) muscles and control processes generating muscle recruitment and forces. Three different impairments of the erector spinae muscles were considered and their actions over 12 consecutive cycles representing a span of 12 months were analyzed. FINDINGS: When asymmetrical muscle degeneration was simulated and weaker erector spinae muscles were located on the convex side of the curve, mild scoliosis (Cobb angle of 8-19 degrees ) was induced in the frontal plane and the kyphosis increased from 72 degrees to 110 degrees in all simulations. Those changes were accompanied by a substantial increase of muscle activity in the Rectus Abdominus and Obliquus Internus. INTERPRETATION: Scoliosis as documented in the literature were induced through an asymmetrical activity in the erector spinae muscles and it can be hypothesized that the Rectus Abdominus and Obliquus Internus have a role in maintaining balance and counteracting against spine torsion. This study demonstrated the feasibility of the modeling approach to investigate a musculo-skeletal deformation process based on a neuromuscular deficit.

The staircase phenomenon at the corrugator supercilii muscle in comparison with the hand muscles.

BACKGROUND: Phonomyography (PMG) is a novel method to monitor neuromuscular block. It is non-invasive and can be applied to any muscle. It can be used interchangeably with mechanomyography (MMG). The staircase phenomenon has not been investigated for this method or at the corrugator supercilii muscle. The purpose of this work was to determine the staircase effect at three different muscles using two different methods. METHODS: In 10 patients undergoing general anaesthesia with sevoflurane, using a laryngeal mask airway without the aid of neuromuscular block, one piezo-electric microphone each was applied to the corrugator supercilii muscle and the first dorsal interosseus muscle. In addition, a force transducer was attached to the tip of the thumb to determine the force of the adductor pollicis muscle. Supramaximal stimulation at 1 Hz was used at the ulnar and the facial nerve. All signals were simultaneously recorded for 30 min. Data are presented as means (SD). RESULTS: The staircase effect was significantly positive for the first dorsal interosseus muscle and the adductor pollicis muscle. The signal potentiation was not significantly different between the first dorsal interosseus muscle with a maximum increase at 148 (19)% using PMG, and the adductor pollicis muscle at 154 (22)% using MMG. The evoked signals reached a plateau after 15-18 min at both muscles. There was only a small initial increase in signal height at the corrugator supercilii to a maximum of 117 (20)% at 7 min, after which the signals decreased to reach a plateau at 25 min. In comparison with the signal height of 105 (25)% at 30 min, there was no significant difference of signal heights throughout the observation period. CONCLUSIONS: A positive staircase phenomenon is found equally at the first dorsal interosseus muscle and the adductor pollicis muscle. There is no significant staircase effect at the corrugator supercilii muscle.

Effects of various adjuvants and a viral infection on the antibody specificity toward native or cryptic epitopes of a protein antigen.

An immunization protocol that induces antibodies (Abs) directed to cryptic epitopes of a protein antigen (Ag) reduces the efficacy of vaccines that ideally should induce Abs against native epitopes. We have shown earlier that viral infections concomitant with immunization against a protein tend to shift the Ab specificity toward cryptic epitopes and tend to induce the production of autoantibodies (autoAbs). Here, we show the effects of three adjuvants on the Ab specificity in the absence or presence of a viral infection (lactate dehydrogenase-elevating virus or LDV), with human growth hormone (hGH) being, as before, the protein Ag. Pathogen-free CBA/Ht and BALB/c mice were immunized with hGH in the presence of complete Freund's adjuvant (CFA), monophosphoryl lipid A (MPL) or alum, with the animals being either infected with LDV or not infected with LDV. Conventional and competition enzyme-linked immunosorbent assays (ELISAs) indicated that in noninfected mice, CFA induced higher titres of anti-hGH Ab than did MPL or alum, with the Ab being almost totally directed to cryptic hGH epitopes. Strikingly, CFA plus LDV infection in CBA/Ht mice shifted the specificity of the anti-hGH Ab toward native epitopes, whereas the virus decreased the Ab titre when MPL or alum was used. Our Western blot results showed that 70% of mice immunized with hGH in the presence of any adjuvant produced autoAbs against a variety of tissue Ags. The amount of autoAb and the concentration of Ab to hGH cryptic epitopes did correlate, suggesting a relationship between both kinds of Ab. Significant differences were observed in the various effects of adjuvants and the viral infection between the two mouse strains used in this work.

Electromyogram and kinematic analysis of lateral bending in idiopathic scoliosis patients.

In adolescent idiopathic scoliosis (AIS), surgical planning currently relies on spinal flexibility evaluation using lateral bending radiographs. The aim was to evaluate the feasibility of non-invasive dynamic analysis of trunk kinematics and muscle activity in patients with AIS before surgical correction. During various lateral trunk bending tasks, erector spinae (18 sites) and abdominal (four sites) muscle activity was sampled using surface electrodes in ten AIS patients and in ten controls. Simultaneously, the spatial displacements of infrared emitting diodes located on the trunk were sampled. Parameters considered were the heterolateral-to-homolateral root-mean-square EMG ratios R at each site and total lateral bending and thoracic and lumbar curvature angle courses. Main alterations concerned apical muscle activity during left bending tasks. ANOVA results showed a significant effect of side (p = 2.1 x 10(-9)), EMG recording site (p = 1.9 x 10(-16)), pathology (p = 3.9 x 10(-16)) and task (p = 2.2 x 10(-11)) on R ratios. The R ratio at T10 and L1 for a simple lateral bending task during left bending averaged 4.8 (SD 4.3) and 3.0 (SD 3.1) in AIS patients, and 2.3 (SD 2.8) and 1.3 (SD 0.4) in controls (p = 6.4 x 10(-4) and 2.5 x 10(-3), LSD post hoc). This preliminary study allowed the development of a functional, non-invasive, non-irradiating dynamic tool for pre-operative evaluation in AIS.