Partial Labyrinth Removal Restricted to the Superior Semicircular Canal in Focal Combined Transpetrosal Approach: Description and Illustrative Cases.

BACKGROUND AND OBJECTIVES: The labyrinthine structures obstruct the surgical view of the deep petroclival region in the transpetrosal approach. Historically, labyrinthectomy and removal of all 3 semicircular canals, with resultant deafness, was used in patients with ipsilateral functional hearing deficits to improve access. The advent and systematization of superior and posterior semicircular canal removal (transcrusal approach) with good rates of hearing preservation has allowed a redefinition of the possibility of partial labyrinthectomy in patients without previous hearing deficits. The present manuscript is intended to describe a technical refinement of partial labyrinthectomy during focal combined petrosectomy, offering a customization of the approach through the selective removal of the superior semicircular canal for specific types of tumors. METHODS: The use of the technique is demonstrated through surgical drawings, pictures, and videos. The rationale to indicate this new approach is discussed based on clinical cases. RESULTS: Three illustrative clinical cases (petroclival meningiomas) are demonstrated. Functional hearing on the approach side has been preserved in all of them. CONCLUSION: The focal combined transpetrosal approach associated with the superior semicircular canal resection has been a promising surgical technique in the treatment of selected petroclival tumors. It has the potential to further decrease the risks of postoperative auditory and vestibular dysfunctions associated with labyrinthectomies.

Medial malleolus malunion with secondary tibiotalar and syndesmotic incongruence - two case reports and ''box resection'' for its treatment.

INTRODUCTION: Ankle fractures are among the most common injuries treated by orthopaedic surgeons. Malunion is one of the possible complications, and corrective surgery is often used to avoid long-term disability. However, few studies address medial malleolus malunion and the best revision surgery technique is controversial. We describe a new surgical technique, the "box resection". MATERIAL AND METHODS: We present two cases of medial malleolus malunion with secondary talus lateral translation and syndesmotic incongruence. In order to restore a congruent tibiotalar joint, we performed the described technique. RESULTS: We record significant improvement of AOFAS Score and range of motion in both cases, with resolution of pain symptoms. Postoperative radiographs show a congruent mortise with syndesmosis reduction. CONCLUSION: The box resection allowed immediate talus medial translation to its original position. It is a simple and effective solution, with a good functional outcome.

INTRODUCCIÓN: La consolidación viciosa del maléolo interno es una posible complicación en las fracturas maleolares. Su tratamiento es difícil y controvertido, por el momento no hay una solución estándar. Proponemos una nueva técnica quirúrgica para el tratamiento de estos casos que denominamos "resección en caja". MATERIAL Y MÉTODOS: Presentamos dos casos de consolidación viciosa del maléolo interno, asociados con traslación lateral del astrágalo y incongruencia de la sindesmosis, donde aplicamos la técnica quirúrgica descrita. RESULTADOS: En ambos casos, obtuvimos una mejora significativa en el puntaje AOFAS y en el rango de movilidad, así como en la resolución de las molestias por dolor. Las radiografías muestran mortajas congruentes y anatómicas, con una reducción satisfactoria de la sindesmosis. CONCLUSIÓN: La técnica de "resección en caja" es simple y eficaz produciendo un buen resultado clínico y funcional.

Medial malleolus malunion with secondary tibiotalar and syndesmotic incongruence - two case reports and «box resection¼ for its treatment / Mal unión del maléolo medial con incongruencia tibiotalar y sindesmótica: «resección en caja¼ para su tratamiento

Abstract: Introduction: Ankle fractures are among the most common injuries treated by orthopaedic surgeons. Malunion is one of the possible complications, and corrective surgery is often used to avoid long-term disability. However, few studies address medial malleolus malunion and the best revision surgery technique is controversial. We describe a new surgical technique, the «box resection¼. Material and methods: We present two cases of medial malleolus malunion with secondary talus lateral translation and syndesmotic incongruence. In order to restore a congruent tibiotalar joint, we performed the described technique. Results: We record significant improvement of AOFAS Score and range of motion in both cases, with resolution of pain symptoms. Postoperative radiographs show a congruent mortise with syndesmosis reduction. Conclusion: The box resection allowed immediate talus medial translation to its original position. It is a simple and effective solution, with a good functional outcome.

Resumen: Introducción: La consolidación viciosa del maléolo interno es una posible complicación en las fracturas maleolares. Su tratamiento es difícil y controvertido, por el momento no hay una solución estándar. Proponemos una nueva técnica quirúrgica para el tratamiento de estos casos que denominamos «resección en caja¼. Material y métodos: Presentamos dos casos de consolidación viciosa del maléolo interno, asociados con traslación lateral del astrágalo y incongruencia de la sindesmosis, donde aplicamos la técnica quirúrgica descrita. Resultados: En ambos casos, obtuvimos una mejora significativa en el puntaje AOFAS y en el rango de movilidad, así como en la resolución de las molestias por dolor. Las radiografías muestran mortajas congruentes y anatómicas, con una reducción satisfactoria de la sindesmosis. Conclusión: La técnica de «resección en caja¼ es simple y eficaz produciendo un buen resultado clínico y funcional.

Hearing preservation middle fossa approach for intracanalicular vestibular schwannoma in a NF2 patient.

Hearing preservation is a cornerstone in the management of intracanalicular vestibular schwannomas. This video demonstrates a middle fossa approach to an intracanalicular schwannoma and highlights some technical and anatomical nuances relevant to the procedure. The patient had sustained hearing preservation in the postoperative period. There are potential benefits in favor of the middle fossa when the tumor reaches the fundus of the internal auditory canal, but the surgeon's individual experience plays a decisive role in the choice of approach. The video can be found here: https://stream.cadmore.media/r10.3171/2021.7.FOCVID21121.

Tentorial Peeling: Surgical Extradural Navigation to Protect the Temporal Lobe in the Focused Combined Transpetrosal Approach.

BACKGROUND: Transpetrosal approaches have been used for treatment of tumors in the petroclival region for many years. Injury to the temporal lobe, however, has been a potential drawback of the techniques described to date. OBJECTIVE: To describe modifications of the transpetrosal surgical technique, which allows extradural manipulation of the temporal lobe during the focused combined transpetrosal approach. This extra layer of protection avoids mechanical brain retraction, direct trauma to the temporal lobe and disruption of the local venous structures. METHODS: The present manuscript describes an innovative technical nuance based on the combination of the focused combined transpetrosal approach, the peeling of the dural layers of the tentorium, and the reverse peeling of the middle fossa dura mater. Ample illustrative material is provided and illustrative cases are presented. CONCLUSION: Peeling of the dural layers of the tentorium is a promising modification of the transpetrosal approach to increase the safety of the temporal lobe manipulation.

Cosmetic benefits of a novel biomimetic lamellar formulation containing niacinamide in healthy females with oily, blemish-prone skin in a randomized proof-of-concept study.

OBJECTIVE: A randomized study was designed to evaluate the potential cosmetic benefit of a biomimetic, niacinamide-containing moisturizing cream in oily, blemish-prone skin. METHODS: Healthy adult women with oily, blemish-prone skin were randomized to one of three treatment groups: test, control, or positive control. In the test group, subjects used the test product (containing 4% niacinamide), plus the standard cleanser (Simple® Kind to Skin Moisturizing Facial Wash). In the control group, subjects received no moisturizer but used the standard cleanser. In the positive control group, subjects used Vivatinell Acnecinamide® Gel Cream (containing 4% niacinamide) as a moisturizer and Neutrogena Visibly Clear® Spot Clearing Facial Wash (containing 2% salicylic acid) as a cleanser. The positive control regimen was included to provide a comparison for estimates of effect size. The primary objective was to evaluate skin moisturization as a change from baseline in corneometer values at 8 h for the test regimen vs. the control regimen. Analysis of covariance was applied for the primary efficacy analysis. RESULTS: A total of 132 subjects were randomized with 44 included in each treatment group. A significant difference was observed in the primary endpoint for the test regimen compared with the control regimen (least-squares mean difference [95% CI]: 3.12 [0.68, 5.56], P = 0.0128). A trend was observed in favour of the positive control regimen compared with the control regimen. Secondary measurements of moisturization supported the primary efficacy outcome. Assessment of blemishes showed a significant difference between the test regimen vs. the control regimen for change from baseline in mean total blemish count at Week 8 (least-squares mean difference [95% CI]: -1.80 [-3.41, -0.19], P = 0.0290). No statistical comparisons between the positive control group and the test group were performed. CONCLUSION: This study provides proof-of-concept evidence that a novel lamellar lipid moisturizer containing niacinamide, in combination with a standard cleanser, can help moisturize the skin and provide an overall improvement in the complexion appearance of people with blemish-prone skin. STUDY REGISTRATION: NCT03093181.

OBJECTIF: Une étude randomisée a été conçue pour évaluer le bénéfice cosmétique potentiel d'une crème hydratante biomimétique contenant du niacinamide sur une peau grasse sujette aux imperfections. MÉTHODES: Des femmes adultes en bonne santé, à peau grasse sujette aux imperfections, ont été randomisées dans l'un des trois groupes de traitement : test, témoin ou témoin positif. Dans le groupe test, les sujets ont utilisé le produit testé (contenant 4 % de niacinamide), plus le nettoyant standard (Nettoyant visage Simple® doux pour la peau). Dans le groupe témoin, les sujets n'ont reçu aucune crème hydratante mais ont utilisé le nettoyant standard. Dans le groupe témoin positif, les sujets ont utilisé le gel crème Vivatinell Acnecinamide® (contenant 4 % de niacinamide) comme crème hydratante et le nettoyant visage pour réduire les imperfections Neutrogena Visibly Clear® (contenant 2 % d'acide salicylique) comme nettoyant. Le schéma de traitement du groupe témoin positif était inclus pour fournir une comparaison des estimations de la taille de l'effet. L'objectif principal était d'évaluer l'hydratation de la peau par le changement par rapport à la référence des valeurs du cornéomètre à 8 h pour le schéma de traitement testé par rapport au schéma de traitement témoin. Une analyse de covariance a été appliquée pour l'analyse de l'efficacité primaire. RÉSULTATS: Un total de 132 sujets ont été randomisés, dont 44 inclus dans chaque groupe de traitement. Une différence significative a été observée dans le critère d'évaluation principal en faveur du schéma de traitement testé par rapport au schéma de traitement témoin (différence moyenne des moindres carrés [IC à 95 %] : 3,12 [0,68, 5,56], P = 0,0128). Une tendance a été observée en faveur du schéma de traitement témoin positif par rapport au schéma de traitement témoin. Les mesures secondaires de l'hydratation ont appuyé le résultat principal d'efficacité. L'évaluation des imperfections a montré une différence significative entre le schéma de traitement testé par rapport au schéma de traitement témoin en ce qui concerne le changement par rapport à la référence dans le nombre moyen total d'imperfections à la semaine 8 (différence moyenne des moindres carrés [IC à 95 %] : _1,80 [_3,41, _0,19], P = 0,0290). Aucune comparaison statistique entre le groupe témoin positif et le groupe test n'a été réalisée. CONCLUSION: Cette étude fournit des éléments de preuve de concept qu'une nouvelle crème hydratante lipidique lamellaire à base de niacinamide, en association avec un nettoyant standard, peut permettre d'hydrater la peau et fournir une amélioration globale de l'aspect du teint chez des personnes dont la peau est sujette aux imperfections. Numéro d'enregistrement de l'étude : NCT03093181.

Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma.

Flow cytometry has become a highly valuable method to monitor minimal residual disease (MRD) and evaluate the depth of complete response (CR) in bone marrow (BM) of multiple myeloma (MM) after therapy. However, current flow-MRD has lower sensitivity than molecular methods and lacks standardization. Here we report on a novel next generation flow (NGF) approach for highly sensitive and standardized MRD detection in MM. An optimized 2-tube 8-color antibody panel was constructed in five cycles of design-evaluation-redesign. In addition, a bulk-lysis procedure was established for acquisition of ⩾107 cells/sample, and novel software tools were constructed for automatic plasma cell gating. Multicenter evaluation of 110 follow-up BM from MM patients in very good partial response (VGPR) or CR showed a higher sensitivity for NGF-MRD vs conventional 8-color flow-MRD -MRD-positive rate of 47 vs 34% (P=0.003)-. Thus, 25% of patients classified as MRD-negative by conventional 8-color flow were MRD-positive by NGF, translating into a significantly longer progression-free survival for MRD-negative vs MRD-positive CR patients by NGF (75% progression-free survival not reached vs 7 months; P=0.02). This study establishes EuroFlow-based NGF as a highly sensitive, fully standardized approach for MRD detection in MM which overcomes the major limitations of conventional flow-MRD methods and is ready for implementation in routine diagnostics.

Quasi-steady aerodynamic model of clap-and-fling flapping MAV and validation using free-flight data.

Flapping-wing aerodynamic models that are accurate, computationally efficient and physically meaningful, are challenging to obtain. Such models are essential to design flapping-wing micro air vehicles and to develop advanced controllers enhancing the autonomy of such vehicles. In this work, a phenomenological model is developed for the time-resolved aerodynamic forces on clap-and-fling ornithopters. The model is based on quasi-steady theory and accounts for inertial, circulatory, added mass and viscous forces. It extends existing quasi-steady approaches by: including a fling circulation factor to account for unsteady wing-wing interaction, considering real platform-specific wing kinematics and different flight regimes. The model parameters are estimated from wind tunnel measurements conducted on a real test platform. Comparison to wind tunnel data shows that the model predicts the lift forces on the test platform accurately, and accounts for wing-wing interaction effectively. Additionally, validation tests with real free-flight data show that lift forces can be predicted with considerable accuracy in different flight regimes. The complete parameter-varying model represents a wide range of flight conditions, is computationally simple, physically meaningful and requires few measurements. It is therefore potentially useful for both control design and preliminary conceptual studies for developing new platforms.

New insight on the underdrawing of 16th Flemish-Portuguese easel paintings by combined surface analysis and microanalytical techniques.

This paper focusses on the study of the underdrawings of 16th century easel paintings attributed to the workshop of the Portuguese-Flemish Master Frei Carlos. This investigation encompasses multidisciplinary research that relates the results of surface exams (infrared reflectography, standard light photography and infrared photography) with analytical investigations. The surface analysis of Frei Carlos' underdrawings by infrared reflectography has shown heterogeneous work, revealing two different situations: (1) an abundant and expressive underdrawing, revealing a Flemish influence and (2) a simple and outlined underdrawing. This preliminary research raised an important question related to this Portuguese-Flemish workshop and to the analytical approach: Is the underdrawing's heterogeneity, as observed in the reflectograms, related to different artists or is this rather an effect that is produced due to the use of different materials in the underdrawing's execution? Consequently, if different materials were used, how can we have access to the hidden underdrawings? In order to understand the reasons for this dissemblance, chemical analysis of micro-samples collected in underdrawing areas and representing both situations were carried out by optical microscopy, micro Fourier transform infrared spectroscopy (µ-FTIR), scanning electron microscopy coupled with energy dispersive X-ray spectrometry (SEM-EDX) and micro-Raman spectroscopy (µ-Raman). Taking into account the different possibilities and practical and theoretical limitations of surface and punctual examinations in the study of easel painting underdrawings, the methodology of research was adjusted, sometimes resulting in a re-analysis of experimental results. This research shows the importance of combining multispectral surface exams and chemical analysis in the understanding of the artistic creative processes of 16th century easel paintings.

Evaluation of Pb (II) biosorption utilizing sugarcane bagasse colonized by Basidiomycetes.

The contamination of water resources by metallic ions is a serious risk to public health and the environment. Therefore, a great emphasis has been given to alternative biosorption methods that are based on the retention of aqueous-solution pollutants; in the last decades, several agricultural residues have been explored as low-cost adsorbent. In this study, the ability of Pb (II) biosorption using sugarcane bagasse modified by different fungal species was evaluated. The presence of carbonyl, hydroxyl, and carboxyl groups in the biosorbent was observed by spectroscopy in the infrared region. By scanning electron microscopy, changes in the morphology of modified material surfaces were observed. The highest adsorption capacity occurred at pH 5.0, while the shorter adsorbate-adsorbent equilibrium was at 20 min, and the system followed the pseudo-second-order model. The maximum biosorption in isotherms was found at 58.34 mg g(-1) for modified residue by Pleurotus ostreatus U2-11, and the system followed the Langmuir isotherm. The biosorption process was energetically spontaneous with low desorption values. This modification showed great potential for filters to remove Pb (II) and provide the preservation of water resources and animal health.

SPF and UVA-PF sunscreen evaluation: are there good correlations among results obtained in vivo, in vitro and in a theoretical Sunscreen Simulator? A real-life exercise.

OBJECTIVE: Strategies to optimize the development of sunscreens include the use of theoretical sunscreen simulators to predict sun protection factor (SPF) and UVA protection factor (UVA-PF) and in vitro measurements of UVA-PF. The aims of this study were to assess the correlations between (1) SPF and UVA-PF results obtained in a theoretical sunscreen simulator with those observed in vivo (SPF and UVA-PF) and in vitro (UVA-PF) and (2) the results of UVA-PF observed in vitro and in vivo for products in different galenic forms containing or not pigments. METHODS: BASF Sunscreen Simulator software was used to evaluate the theoretical performance of formulations regarding SPF and UVA protection. In vitroUVA-PF and in vivoSPF were determined for all formulations. UVA-PFin vivo measurements were carried out only on products for which the galenic forms (compact foundations and lip balms) or the presence of dye or pigments could make the results of UVA-PFin vitro less reliable (due to a possible uneven film formation). RESULTS: The results of the SPF calculated by the BASF Sunscreen Simulator presented a very good correlation with SPF observed in vivo in the absence of pigments (r = 0.91; P < 0.05) and a good correlation in the presence of pigments (r = 0.70; P < 0.05). The UVA-PF calculated by the BASF Sunscreen Simulator also exhibited a very good correlation with UVA-PF measured in vitro (r = 0.88; P < 0.05) for the formulations not containing pigment and a good correlation (r = 0.75; P < 0.05) for the formulations containing pigment. The correlation of same UVA-PF calculated by BASF Sunscreen Simulator with UVA-PF measured in vivo for the formulations containing pigment was r = 0.74 (P < 0.05), which is considered good. In addition, the measurements of UVA-PFin vivo presented a good correlation with the values obtained in vitro (r = 0.74; P < 0.05). CONCLUSION: In the present study, the use of BASF Sunscreen Simulator and in vitroUVA tests showed good correlations with in vivo results and could be considered as valuable resources in the development of sunscreens.

Error analysis and assessment of unsteady forces acting on a flapping wing micro air vehicle: free flight versus wind-tunnel experimental methods.

An accurate knowledge of the unsteady aerodynamic forces acting on a bio-inspired, flapping-wing micro air vehicle (FWMAV) is crucial in the design development and optimization cycle. Two different types of experimental approaches are often used: determination of forces from position data obtained from external optical tracking during free flight, or direct measurements of forces by attaching the FWMAV to a force transducer in a wind-tunnel. This study compares the quality of the forces obtained from both methods as applied to a 17.4 gram FWMAV capable of controlled flight. A comprehensive analysis of various error sources is performed. The effects of different factors, e.g., measurement errors, error propagation, numerical differentiation, filtering frequency selection, and structural eigenmode interference, are assessed. For the forces obtained from free flight experiments it is shown that a data acquisition frequency below 200 Hz and an accuracy in the position measurements lower than ± 0.2 mm may considerably hinder determination of the unsteady forces. In general, the force component parallel to the fuselage determined by the two methods compares well for identical flight conditions; however, a significant difference was observed for the forces along the stroke plane of the wings. This was found to originate from the restrictions applied by the clamp to the dynamic oscillations observed in free flight and from the structural resonance of the clamped FWMAV structure, which generates loads that cannot be distinguished from the external forces. Furthermore, the clamping position was found to have a pronounced influence on the eigenmodes of the structure, and this effect should be taken into account for accurate force measurements.

Imaging of the cavernous sinus lesions.

This educational paper reviews the normal anatomy of the cavernous sinus (CS) and the imaging findings of common and uncommon lesions of this region. CS lesions may arise from different components of the CS or from adjacent structures and spaces. They can be classified as tumoral, inflammatory/infectious, vascular and congenital. Tumoral lesions include benign (meningiomas, pituitary adenomas, schwannomas) and malignant neoplasms (chondrosarcomas, chordomas, nasopharyngeal carcinomas, leukemia, metastases). Inflammatory/infectious conditions comprise: Tolosa Hunt, abscess, Lemierre syndrome and thrombophlebitis. Vascular lesions include: hemangiomas, carotido-cavernous fistula, aneurysms, arteriovenous malformations. Congenital conditions include the epidermoid cyst, dermoid cyst and fatty deposits. Although imaging features of non-vascular CS diseases are most often non-specific, careful analysis of the adjacent structures suggests the correct diagnosis. In vascular pathology, characteristic MR imaging findings are observed.

Induction of reversible meiosis arrest of bovine oocytes using a two-step procedure under defined and nondefined conditions.

The objective was to study the effect of a defined culture system, on nuclear and cytoplasmic maturation of bovine oocytes, using the two-step procedure of IVM to detect possible inhibition and subsequent resumption of meiosis arrest. In the first step, called the prematuration period (PMP), COCs were cultured in T1-non-defined medium (NDM), or T2-defined medium (DM), both for 24 h. In step 2, called the resumption period (RP), COCs were cultured in: NDM (T1); DM + NDM (T3); or DM+DM (T4) for 24 h in each medium. The NDM was composed of TCM-199 supplemented with FCS and FSH. The DM was composed of alpha-MEM supplemented with PVA, insulin, IGF-1, androstenedione, nonessential amino acids, transferrin, and sodium selenium. Oocytes from T2 had a lower (P < 0.05) rate of nuclear maturation (19.8%) than T1 oocytes (83.2%). Also, T2 COCs appeared to be in the process of cytoplasmic maturation, according to the distribution of organelles assessed by transmission electron microscopy (MET). These COCs had characteristics previously described as mature: erect microvilli on the plasmembrane, presence of cortical/evenly distributed mitochondria throughout the ooplasm, and presence of 50% aligned/cluster cortical granules. Immature characteristics such as small PvS, compact cumulus cells, and presence of 50% cortical granule clusters were also observed. The T1 COCs had only characteristics of maturation (P < 0.05). In step 2 (RP), meiosis arrest induced by DM was resumed after an additional 24 h of culture in NDM (T3) with 79.2% mature COCs, whereas in T4, meiosis arrest was maintained, resulting in almost 70% immature COCs (P < 0.05). At the end of RP, T3 COCs had the mature characteristics of mitochondria spread throughout the cytoplasm (P < 0.05), cumulus expansion, and alignment of cortical granules, whereas the T4 group had both immature and mature characteristics. We inferred that DM can be used in lieu of meiosis inhibitors and furthermore, it can provide extra time to study nuclear and cytoplasmic maturation synchrony of IVM.

Assessment of chemically modified sugarcane bagasse for lead adsorption from aqueous medium.

This study evaluated the adsorption capacity of chemically modified sugarcane bagasse with citric acid (B-CA), sodium hydroxide (B-S) and citric acid and sodium hydroxide (B-CAS) for Pb2+ ion adsorption in aqueous medium. Infrared spectroscopy (FT-IR) was used to characterise the materials, where the chemical modification was confirmed by the presence of carboxylate groups created at 1,730 cm(-1) peak. All assays related to Pb2+ ion adsorption onto adsorbent, i.e. equilibrium time between Pb2+ ions and adsorbate (24 h), as well as Pb2+ ion concentration, were performed in batch system. The initial and final Pb2+ ion concentration after shaking time was determined by Flame Atomic Absorption Spectrometry (FAAS). Isotherm adsorptions were applied to Langmuir and Freundlich linear models and maximum adsorption capacity (MAC) of materials towards Pb2+ ions was calculated in function of modifications. A significant improvement regarding Pb2+ ion adsorption after the bagasse treatment with citric acid, in which MAC was 52.63 mg g(-1), was observed. The adsorptions followed the behaviour described by the Langmuir linear model and its kinetics follow the behaviour described by the pseudo-second-order equation.

Influence of human leucocyte antigen-DRB1 on the susceptibility to rheumatoid arthritis and on the production of anti-cyclic citrullinated peptide antibodies in a Portuguese population.

OBJECTIVE: To clarify the influence of the HLA-DRB1 locus on the susceptibility to rheumatoid arthritis and the production of anti-cyclic citrullinated peptide antibodies (anti-CCP) in a Portuguese population. METHODS: 141 patients with rheumatoid arthritis fulfilling the American College of Rheumatology 1987 revised criteria for rheumatoid arthritis were compared with 150 healthy controls. Human leucocyte antigen (HLA)-DRB1 locus genotyping was assessed by polymerase chain reaction reverse probing assays and sequence-specific primers. Anti-CCP antibodies were quantified by ELISA in patients with rheumatoid arthritis. Frequencies between groups were compared by the two-sided Fisher's exact test and considered significant if p<0.05. RESULTS: The HLA-DRB1*04 and HLA-DRB1*10 groups were highly associated with rheumatoid arthritis (p<0.001 and p = 0.031, respectively). High titres of anti-CCP antibodies were largely associated with the presence of HLA-DRB1*04/10. CONCLUSION: The well-recognised susceptibility alleles to rheumatoid arthritis, HLA-DRB1*04, were associated with rheumatoid arthritis in Portuguese patients. The relatively rare DRB1*10 was also associated with rheumatoid arthritis, as was described previously in other southern European countries. Both groups were associated with high anti-CCP titres, reinforcing its relevance to disease onset.

Multiplexed immunoassay for detection of rheumatoid factors by FIDISTM technology.

Our objective was to (i) compare FIDIS Rheuma, a new multiplexed immunoassay designed for simultaneous detection of IgM class rheumatoid factors (RF) directed against Fc determinants of IgG from humans and animals, with agglutination and ELISA (conventional methods) and (ii) evaluate the clinical sensitivity and specificity of biological markers for rheumatoid arthritis (RA). To do this, FIDIS technology was employed using the Luminex system. It consists of distinct color-coded microsphere sets, a flow cytometer, and digital signal processing hardware and software. Agglutination and ELISA tests were performed with commercial kits. The study included 134 samples from RA patients and 105 from healthy blood donors. For human specificity, we compared FIDIS with latex agglutination and ELISA. Relative sensitivities were 98.9% and 88.5% and specificities were 90.2% and 94.6%, respectively. For animal specificity, we compared FIDIS with Waaler-Rose and ELISA. The results were 84.9% and 71.9% for the sensitivities and 97.5% and 98.4% for the specificities, respectively. Detection of IgG anti-CCP by ELISA and IgG antikeratin by immunofluorescence was also determined in order to compare their clinical sensitivity and specificity with IgM-RF, according to the method used. The results were: IgG anti-CCP 72.3%, 97.2%; IgG antikeratin 36.6%, 100%; latex agglutination 66.4%, 97.2%; Waaler-Rose 55.9%, 96.3%; FIDIS human 73.9%, 92.1%; FIDIS animal 49.2%, 97.2%; ELISA human 93.2%, 95.5%; and ELISA animal 74.6%, 91.3%. The results showed the efficiency of FIDIS with analytical performance equivalent to the conventional methods, but having the advantage of giving quantitative results (IU/mL).

Detection of hepatitis B virus core mutants by PCR-RFLP in chronically infected patients.

HBV chronic infection is an important health problem. The HBV core antigen carries several epitopes for T and B cell recognition and the immune response is crucial for determining the outcome of viral infection. Using PCR-RFLP several point mutations were detected in the HBV core ORF of HBV extracted from the serum of 140 chronically infected patients and 86 samples from another 37 patients followed-up in a longitudinal study. Mutations at position 2248 and 2147 (A3) and at 2038 (M2) were found most frequently. The wild type core genotype was found in about 50% of the samples. PCR-RFLP results were confirmed by direct sequencing of amplified products from HBV DNA present in chronically infected patients. The method is rapid and reliable and may be particularly useful for a rapid detection of viral mutants in a large number of patients.

Cross-reactivity between Chlamydia trachomatis heat shock protein 10 and early pregnancy factor.

Chlamydia trachomatis heat shock protein 10 (Chsp10) is associated with chronic genital tract infection with C. trachomatis. Chsp10 is homologous to human chaperonin 10 (Cpn10) and early pregnancy factor (EPF), a form of human Cpn10 that is specifically secreted at the start of pregnancy. We investigated cross-reactions between serum anti-Chsp10 antibodies and anti-EPF antibodies in pregnant and nonpregnant patients. Pregnancy was found to be associated with the presence of anti-EPF antibodies, which are specifically induced in pregnant women with a history of C. trachomatis infection, and with the presence of serum anti-Chsp10 antibodies. We also found that infertility was associated with the presence of anti-Chsp10 and anti-EPF antibodies. The HLA class II haplotype DR8 DQ4 was associated with the presence of anti-Chsp10 antibodies but not of anti-EPF antibodies.

Filtration induces changes in activity states and leucocyte populations.

The distribution of leucocyte subpopulations in platelet concentrates (PC) derived from pre-storage filtered platelet-rich plasma (PRP), the cell suspension obtained by reverse filter washing and the post-filtered PC, were monitored by immunophenotyping analysis using CD3, CD20 and CD33. Leucocyte activation analysis with the CD11b marker revealed that this molecule is up regulated in neutrophils taken from the filter. This, together with the loss of cell viability during the enrichment process, suggests that contact with the filter matrix and processing and storage of samples containing leucocytes may lead to activation and loss of leucocyte viability. These changes were found to be more pronounced in less stable myeloid cells and account for the differences reported among various authors which in some cases related to operational conditions such as the enrichment process used and the length of time between filtration and analysis of samples. Finally, statistical analysis of the results obtained by immunophenotypic studies indicate that post-filter samples (S) contain significantly higher numbers of CD33+ myeloid cells when compared to (PF) the pre-filter samples (65.03%+/-12.6 and 24.56%+/-14.73, p<0.0000), with a decrease in T cells (50.72%+/-14.80 in PF and 24.05+/-9.48 in the cell suspension (S), p<0.0007) and B cells (14.96+/-9.31 in PF and 9.9+/-5.22 in S, p<0.201). A new strategy for assessing the influence of the filtration process on residual leucocyte activation and viability is described. This has direct relevance to collection, processing, storage and quality monitoring of PC.