Search

The integration of Building Information Modelling (BIM) and Geographical Information Systems (GIS) is gaining momentum in digital built Asset Management (AM), and has the potential to improve information management operations and provide advantages in process control and delivery of quality AM services, along with underlying data management benefits through entire life cycle of an asset. Work has been carried out relating GeoBIM/AM to buildings as well as infrastructure assets, where the potential financial savings are extensive. While information form BIM maybe be sufficient for building-AM; for infrastructure AM a combination of GIS and BIM is required. Scientific literature relating to this topic has been growing in recent years and has now reached a point where a systematic analysis of current and potential uses of GeoBIM in AM for Infrastructure is possible. Three specific areas form part of the analysis – a review of BIM and Infrastructure AM and GIS and Infrastructure AM leads to a better understanding of current practice. Combining the two, a review of GeoBIM and Infrastructure AM allows the benefits of, and issues relating to, GeoBIM to be clearly identified, both at technical and operational levels. A set of 54 journal articles was selected for in-depth contents analysis according to the AM function addressed and the managed asset class. The analysis enabled the identification of three categories of issues and opportunities: data management, interoperability and integration and AM process and service management. The identified knowledge gaps, in turn, underpin problem definition for the next phases of research into GeoBIM for infrastructure AM.

Table of contents S1 Criteria for exercises selection in subjects with low back pain Alessandro Aina S2 Recent advances in pathophysiology and treatment of myofascial trigger points Marco Barbero S3 Rehabilitation of scapular dyskinesia Barbara Cagnie S4 Musculoskeletal rehabilitation in subjects affected by neurological disorders Elena Castelli S5 Which examination tests suggest the best candidates for manual therapy Chad Cook S6 Case study: the role of the measurements for the identification of targets and guidance of the treatment Silvano Ferrari S7 Assessment of joint mobility: state of the art Andrea Foglia, Paolo Bizzarri S8 Core stabilization exercises in the treatment of urinary incontinence Donatella Giraudo S9 Preventing surgical subacromial decompression through rotator cuff rehabilitation Chris Littlewood S10 Methodological aspects of Clinical Prediction Rules in the rehabilitation of Low Back Pain Paolo Pillastrini S11 Interpretability of outcome measures in musculoskeletal rehabilitation Daniele Piscitelli S12 Conservative treatment of the misalignment of the spine: state of the art and perspectives Michele Romano S13 Balance training in subjects with musculoskeletal disorders Andrea Tettamanti S14 Dosage of manual therapy: principles for clinical practice Carla Vanti S15 Are there speed limits in post-surgery lower limb rehabilitation? Stefano Vercelli S16 Classification of predominant neuropathic, nociceptive or central sensation pain Lennard Voogt P1 A wearable proprioceptive stabilizer (Equistasi®) for rehabilitation of balance disorders in multiple sclerosis patients: preliminary results of a randomized, double-blind, versus placebo controlled study Aceto Maria, Spina Emanuele, Paone Paolo, Silvestre Francesco, Carotenuto Antonio, Cerillo Ilaria, Orefice Giuseppe P2 Effect of repeated neck retraction movements on strength and EMG activity of the upper limbs, range of motion and cervical posture Bassi Raffaele, Fiorito Serena, Aina Alessandro P3 Hamstring injuries: clinical assessment or image evaluation? Bonfanti M., Pasquetti M. P4 Effectiveness of the physical therapy treatment on pelvic floor muscles in pelvic girdle pain. A literature review Bortolami Arianna, Pillastrini Paolo, Vanti Carla P5 Scapulo-humeral muscles electromiographic activity during the elevation movement of the upper limb in subjects with rotator cuff lesion Brioschi D, Vitali M, Pedretti A, Fraschini G, Tettamanti A P6 Mechanical low back pain: secular trend and intervention topics of randomized controlled trials Castellini G, Gianola S, Bonovas S, Banfi G, Moja L P7 Quality of reporting in rehabilitation interventions for low back pain: a review of published randomised controlled trials Greta Castellini, Silvia Gianola, Pamela Frigerio, Michela Agostini, Rosa Bolotta, Davide Corbetta, Monica Gasparini, Paolo Gozzer, Erica Guariento, Linda Li, Valentina Pecoraro, Valeria Sirtori, Andrea Turolla, Andreano A, Lorenzo Moja P8 Power analysis and sample size reporting in rehabilitation of low back pain: review of randomized controlled trials included in Cochrane systematic review Castellini G, Gianola S, Bonovas S, Moja L P9 A core outcome set for clinical trials in non-specific low back pain Alessandro Chiarotto, Caroline B. Terwee, Maarten Boers, Raymond W. Ostelo P10 Roland & Morris Disability Questionnaire and Oswestry Disability Index: which has better measurement properties? A systematic review and meta-analysis Alessandro Chiarotto, Lara J. Maxwell, Caroline B. Terwee, George A. Wells, Peter Tugwell, Raymond W. Ostelo P11 Prevalence of myofascial trigger points in spinal pain disorders: systematic review and meta-analysis Alessandro Chiarotto, Ron Clijsen, Cesar Fernandez-de-las-Penas, Marco Barbero P12 Post-surgical scar rating scales in physiotherapy: a systematic review Ciceri Matteo, Rossetti Sara, Vercelli Stefano P13 Efficacy of action observation pre-operative training in functional recovery after hip and knee prosthesis Cislaghi M, Penone G, Marinelli G, Rezzan G, Melegati G, Gatti R P14 The use of Kinesio Taping in the treatment of hematomas: can we distinguish fancy from the effects? Colombo Claudio, Tolosa Francesca, Andrea Moriondo, Vercelli Stefano P15 Who is able to perform explicit motor imagery after stroke? Stefano Doronzio, Matteo Paci P16 Responsiveness of the Bridge tests in Symptomatic Lumbar Spondylolisthesis Silvano Ferrari, Carla Vanti, Marco Monticone P17 The relationship between number of sessions and clinical results in lumbar symptomatic spondylolisthesis Silvano Ferrari, Carla Vanti, Marco Monticone P18 Conservative treatments for upper extremity tendinopathies in occupational medicine: narrative review Garzonio Fabiola, Zanetta Anna, Bargeri Serena, Cerone Giorgia, Sartorio Francesco P19 The time-related effect of roller-massager on extensibility of the hamstring muscles Filippo Ghirlanda, Alessandro Schneebeli, Corrado Cescon, Marco Barbero P20 Effect of two different exercise programs on pain, disability and quality of life in people with subacute and chronic nonspecific neck pain Gioia G, Faccendini S, Aina A, Tettamanti A P21 A bridge between clinical practice and research: how health professionals can study literature and learn on the job Granzotto G, Coppola L, Gava I, Frassinelli M, Gattinoni F P22 Walking capacity improves after neuromotor physiotherapy in stroke patients Lorenzo Guidotti, Marco Postiglione, Bruna Lombardi, Matteo Paci P23 Intra and inter-session reliability of the angle between pain onset and submaximal pain during upper limb neurodynamics test 1: a study in healty individuals Diego Leoni, Davide Storer, Roberto Gatti, Michele Egloff, Marco Barbero P24 Effects of an AO-MI training on balance task in patients affected by multiple sclerosis Magno Tiziano, Tettamanti Andrea P25 Outcomes of treatment with neuromuscular stimulator for chronic anterior knee pain: a clinical case Daniele Maremmani, Sebastiano Cencini, Giuseppe Plebani P26 Consistency in locating pressure stimuli over the lumbar spine on a digital body chart: a comparison between chronic low back pain patients and healthy subjects Federica Moresi, Marco Barbero, Matteo Isnardi, Alberto Gallace, Corrado Cescon, Roberto Gatti P27 Malignant Cord Compression in the thoraco-lumbar spine. Early signs and symptoms in the differential diagnosis of low back pain Moretti N, Maselli, Testa M. P28 End growth results of exercise treatment to avoid bracing in adolescents with idiopathic scoliosis: a prospective cohort controlled study Stefano Negrini, Sabrina Donzelli, Francesco Saveri, Alessandra Negrini, Silvana Parzini, Michele Romano, Fabio Zaina P29 Reliability of the Ashworth scale and its modified versions: systematic review and meta-analysis Leonardo Nesi, Francesco Ferrarello, Valeria Anna Maria Bianchi, Matteo Paci P30 Differences in motor recovery between upper and lower limbs in stroke subtypes Matteo Paci, Luca Nannetti, Bruna Lombardi P31 Influence of clinical experience on the reliability of the Salford Gait Tool Giuditta Mini, Mariangela Marchettini, Francesco Ferrarello, Matteo Paci P32 A somatosensory discrimination training induces brain functional changes in healthy young subjects Fabio Piccolo, Federica Agosta, Elisabetta Sarasso, Paola Adamo, Federico Temporiti, Andrea Falini, Roberto Gatti, Massimo Filippi P33 Effects of dynamic tilt-table with integrated robotic stepping associated with functional electrical stimulation: a cross-over study Daniele Piscitelli, Roberto Meroni, Leonardo Pellicciari, Marco A. Mondelli, Thomas Favaron, Cesare G. Cerri, Enrico A. Tallarita P34 Manual therapy techniques in the treatment of whiplash and its associated disorders: a systematic review Ravizzotti Elisa, Vercelli Stefano P35 The Patient and Observer Scar Assessment Scale (POSAS) as a screening tool for early detection of pathologic post-surgical scars in physiotherapy Rossetti Sara, Ciceri Matteo, Vercelli Stefano P36 The anatomical basis of genetic dystonia: a multimodal MRI study Elisabetta Sarasso, Federica Agosta, Aleksandra Tomić, Silvia Basaia, Nataša Dragašević, Marina Svetel, Massimiliano Copetti, Vladimir S. Kostic, Massimo Filippi P37 The relative inclination of the end vertebrae of a scoliotic single curve can influence the results of the conservative treatment? A pilot study Francesco Saveri, Michele Romano, Matteo Mastrantonio, Alessandra Negrini, Fabio Zaina, Negrini Stefano P38 Physiotherapy for myofascial pain syndromes: reported methodological quality of randomized controlled trials indexed in the PEDro database Alessandro Schneebeli, Greta Castellini, Valentina Redaelli, Emiliano Soldini, Marco Barbero P39 Is the patellar pubic percussion test useful to diagnose only femur fractures or something else? Two case reports Segat M, Casonato O, Margelli M, Pillon S. P40 Effectiveness of new rehab method for MDI Spunton V, Fenini R, Garofalo R, Conti M P41 Toe walking and autism: cross-sectional study on clinical presentation patterns and correlation with language delay Valagussa G, Balatti V, Trentin L, Melli S, Norsi M, and Grossi E P42 Active Kyphosis Value (AKV): a new test for the evaluation of the kyphotisation mobility of the thoracic spine Massimiliano Vanossi, Francesco Saveri, Michele Romano P43 Pain Drawing and psychological distress in low back pain - systematic review and meta-analysis Carla Vanti, Sara Taioli, Ivan Gardenghi, Lucia Bertozzi, Anna Rosso, Antonio Romeo, Paolo Pillastrini P44 Responsiveness of the Oswestry Disability Index in symptomatic lumbar spondylolisthesis Carla Vanti, Silvano Ferrari, Martina Ruggeri, Marco Monticone P45 The relationship between instability tests, pain and disability in non-specific low back pain Carla Vanti, Bellini Filippo, Cristina Conti, Federica Faresin, Martina Ruggeri, Raffaella Piccarreta, Silvano Ferrari P46 Smartphone applications (apps) for physical therapists: a review Villanova Luca, Vercelli Stefano P47 A randomized controlled pilot trial of hand robotic training compared with a sensory-motor training program in post stroke patients Violini Claudia, Cenci Marco Joseph, Delconte Carmen, Pisano Fabrizio P48 Critical Illness PolyNeuroMyopathy (CIPNM): chance for a good prognosis Youssef S, Montesano M, Picardi M, De Giampaulis P, Corbo M, Pisani L. P49 The EdUReP approach plus manual therapy for the management of insertional Achilles tendinopathy: a case study Zanetta Anna, Garzonio Fabiola, Ruella Carolina, Sartorio Francesco

Objective: Cognitive impairment is a core feature of schizophrenia, related to dopaminergic dysfunction in the prefrontal cortex (PFC). It is hypothesized that functional single nucleotide polymorphism (SNP) rs4680 of the catechol-O-methyltransferase (COMT) gene could mediate the relationship between cognition and dopamine activity in the PFC. Other COMT SNPs could also play a role. Methods: We evaluated the role of three COMT SNPs (rs737865, rs165599, and rs4680) in schizophrenia and their impact on three working memory tasks. For genetic association analyses, 212 individuals with schizophrenia and 257 healthy controls (HCs) were selected. The Visual Working Memory (VWM) Task, Keep Track Task, and Letter Memory Task were administered to 133 schizophrenics and 93 HCs. Results: We found a significant association of rs737865, with the GG genotype exerting a protective effect and the GA haplotype (rs4680/rs165599) exerting a risk effect for schizophrenia. COMT rs4680 AA carriers and rs737865 AA carriers scored lowest on the Keep Track Task. When the genotype*group interaction effect was evaluated, rs165599 exerted opposite effects for VWM and Keep Track task performance in patients and controls, with AA carriers scoring lowest on both tests among controls, but highest among patients. Conclusion: These data support the hypothesis that COMT polymorphisms may be associated with schizophrenia and modulate cognition in patients and controls.

A partir de la intervención rusa en la crisis de Crimea y la suspensión de las relaciones con la Organización del Tratado del Atlántico Norte (OTAN) en 2014, las imágenes de la guerra fría se volvieron a plasmar con una tensión política y militar en grandes espacios territoriales y marítimos entre ambos actores. Este trabajo busca analizar el posicionamiento geopolítico de la OTAN frente Rusia en el Mar Mediterráneo entre el 2014-2015. Para ello se tomarán como referencia las estrategias relacionadas a la maximización de poder y balance disuasorio entre los actores en cuestión.

Certain monoterpenes produced by aromatic plants are known to have lethal and sublethal effects on insects. As there is a need to replace conventional pesticides (organophosphates, carbamates, pyrethroids) with biorational pesticides, we evaluated the behavioural and toxicological responses of the German cockroach, Blattella germanica, exposed to the following monoterpenes, namely (+)-a-pinene, (-)-a-pinene, limonene, menthone, linalool, menthyl acetate and geraniol. Locomotor activity and repellency were quantified using a video tracking system. To study the effect on locomotor activity, larvae were exposed to filter papers treated with concentrations of monoterpenes ranging between 1.4 and 1,400 µg/cm2. Only menthyl acetate applied at a concentration of 14 µg/cm2 or higher increased the locomotor activity of the larvae. Repellency was evaluated by exposing larvae to circles of filter paper divided in two equal zones: one zone was treated with a solution of a monoterpene in acetone (14 or 140 µg/cm2) and the other with only acetone. Larvae were repelled by all the monoterpenes tested. The two concentrations of geraniol and the highest concentration of menthyl acetate and linalool were as repellent as DEET (positive control). The fumigant effect was evaluated by exposing larvae to vapour from 100 µl of pure monoterpene in a sealed container. The values of the 50% Knock-down Times (KT50), expressed in minutes, were: (+)-a-pinene: 11.8, (-)-a-pinene: 14.6, limonene: 81.0, menthone: 141.0, and linalool: 238.6. (+)-a-Pinene was as good a fumigant as dichlorvos (positive control, KT50 = 9.7 min). The least effective compounds were menthyl acetate and geraniol (both affected less than 50% of larvae after 490 min of exposure). In conclusion, (+)-a-pinene (for its fumigant effect), menthyl acetate (for its hyperactivant effect) and menthone and geraniol (for their repellent properties) could potentially be used for controlling B. germanica.

OBEJTIVO: avaliar o nível de desenvolvimento cognitivo e afetivo-social de crianças com diagnóstico de deficiência auditiva. MÉTODO: participaram do estudo 50 crianças com diagnóstico de deficiência auditiva, com idade média de 16,1 meses, de ambos os gêneros. Empregou-se a entrevista semi-estruturada com os pais para a obtenção da história de vida da criança, a Escala de Desenvolvimento Comportamental e a Escala de Avaliação da Reação de Retração do Bebê. RESULTADO: o estudo identificou 80% das crianças com perda auditiva profunda bilateral. O Quociente de Desenvolvimento na normalidade ocorreu em 76% da amostra, sendo as funções da linguagem e pessoal-social, as mais prejudicadas. Os níveis de ajustamento afetivo e interacional foram caracterizados pelas reações de alarme para problemas de interação pessoal-social. CONCLUSÃO: as competências defasadas de habilidades verbais e não-verbais mostraram implicações no desenvolvimento cognitivo, sugestivas da condição da criança com perda auditiva e dos fatores de risco associados, com limitação circunstancial a trocas comunicativas que promovem o desenvolvimento de competências adaptativas e o fortalecimento da auto-estima para as relações.

Introduction: People with mild haemophilia (PWMH) experience sporadic bleeds and are less likely to receive an early diagnosis, appropriate treatment and medical care. Arthropathy is a key determinant of health-related quality of life (QoL), producing pain, limitations in mobility and daily activities. The aim of this study is to evaluate the incidence, risk factors and QoL associated with arthropathy in PWMH., Materials and Methods: Observational, cross-sectional cohort study. Data were collected in a single interview and evaluated by a physiotherapist and an orthopaedist and analysed on demographics; baseline factor levels; as well as clinical (Haemophilia Joint Health Score [HJHS]), ultrasound (Haemophilia Early Arthropathy Detection with Ultrasound [HEAD-US]), radiological (Pettersson score [PS]), pain (visual analogue scale [VAS]) and QoL evaluations. We defined arthropathy when at least one of the joints shown with a HEAD-US score ≥ 1., Results: Eighty-five patients and 510 joints were included. Patients' mean age was 35.9 years-old. Median age was 44.2 in patients with arthropathy versus 14.9 in patients without; the difference was statistically significant (p < .001). In patients over 20 years old, 90.5% shown arthropathy. Only 24 (28%) patients had no joint damage (HEAD-US = 0), and 61 (72%) had at least one joint with a HEAD-US ≥ 1. The ankle was the most affected joint. Patient age was found to be the most important risk factor associated with the development of arthropathy., Conclusions: Joint damage as a result of prior hemarthrosis was the most relevant factor associated with lower QoL, and emphasised the importance of early diagnosis and appropriate management in this particular population., (© 2023 John Wiley & Sons Ltd.)

Key Message: Managing forest residues according to the carbon content of the soil helps to minimize the ecological footprint of their removal., Context: In Mediterranean mountain ecosystems, unsustainable harvesting of wood residues might contribute to land degradation, carbon, and nutrient depletion in forest soils., Aims: This study aimed to assess the amount of forest biomass residues that should be left on-site to minimize the depletion of soil fertility., Methods: We estimated the availability of biomass residues in the public forest land of the Basilicata region of Southern Italy by collecting stand-scale inventory attributes from forest management plans. Subsequently, we quantified the amount of forest biomass residue released by implementing a scenario-based approach., Results: Approximately 5800 m 3  year -1 of forest residues could be potentially available for bio-based industries at the regional scale within the next 10 years. Such residues mainly belong to broadleaved forest types, having a high variability in their soil organic stock (228.5-705.8 Mg C ha -1 ) and altitudinally spanning from 400 to 1500 m a.s.l. In these forests, the simulated scenarios displayed a wide range of average harvestable residues from 2.5 to 5.5 m 3  ha -1 , containing approximately 1.1 to 2.1 Mg ha -1 of organic carbon., Conclusion: Our study suggests that forest management plans are a useful source of information to estimate the available forest biomass residues consistently. In southern Mediterranean mountain forests, the management of forest residues according to soil carbon content helps to minimize the environmental impact and increase their sustainability., Competing Interests: Competing interestsThe authors declare that they have no conflict of interest., (© The Author(s) 2022.)

Objective: The purpose of this research is to assess the analgesic efficiency of Photobiomodulation (PBM) in pain reduction in young patients during rapid maxillary expansion therapy. Materials and Methods : Thirty patients were included and allocated to an experimental group [Rapid Palatal Expansion (RPE) and PBM] and a control group (RPE only) at random. Inclusion criteria were skeletal age from CVS1 to CVS3, examined on the cephalometric lateral teleradiography, with cervical vertebra staging and completed eruption of upper first molar. Exclusion criteria were previous orthodontic treatment, bone disease, disability, or skeletal age from CVS4. Patients referred the pain they felt using a Numerical scale rate (NRS), ranging from 0 to 10, with specific intervals of time: 6 h, 12 h, 24 h, and from days 2 to 7. The Wilcoxon-Mann-Whitney test was used to evaluate differences in NRS reported values between the two groups. Results: The final sample included 30 patients, 14 males and 16 females, with a mean age of 7.8 ± 1.2 years. The pain that was felt at each time interval and the maximum score of pain were significantly lower in the experimental group ( p < 0.05) and decreased faster in the experimental group, with a score test near to 0 after 2/3 days. Conclusions : PBM is efficient in reducing the intensity and the time of pain felt by young patients that undergo rapid maxillary expansion.

Protein lysine acetylation has emerged as a major regulatory post-translational modification in different organisms, present not only on histone proteins affecting chromatin structure and gene expression but also on nonhistone proteins involved in several cellular processes. The same scenario was observed in protozoan parasites after the description of their acetylomes, indicating that acetylation might regulate crucial biological processes in these parasites. The demonstration that glycolytic enzymes are regulated by acetylation in protozoans shows that this modification might regulate several other processes implicated in parasite survival and adaptation during the life cycle, opening the chance to explore the regulatory acetylation machinery of these parasites as drug targets for new treatment development., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The genetic engineering method CRISPR has been touted as an efficient, inexpensive, easily used, and targeted genetic modification technology that is widely suggested as having the potential to solve many of the problems facing agriculture now and in the future. Like all new technologies, however, it is not without challenges. One of the most difficult challenges to anticipate and detect is gene targets that are inaccessible due to the chromatin state at their specific location. There is currently no way to predict this during the process of designing a sgRNA target, and the only way to detect this issue before spending time and resources on full transformations is to test the cleavage ability of the sgRNA in vivo . In wheat, this is possible using protoplast isolation and PEG transformation with Cas9 ribonucleoprotein complexes. Therefore, we have developed a streamlined protocol for testing the accessibility of sgRNA targets in wheat. The first steps involve digesting wheat leaf tissue in an enzymatic solution and then isolating viable protoplasts using filters and a sucrose gradient. The protoplasts are then transformed using Cas9 ribonucleoprotein complexes via PEG-mediated transformation. DNA is isolated from the CRISPR-Cas-edited protoplasts and PCR is performed to amplify the gene target region. The PCR product is then used to assess the editing efficiency of the chosen sgRNA using Sanger sequencing. This simplified protocol for the isolation and transformation of wheat protoplast cells using Cas9 ribonucleoprotein complexes streamlines CRISPR transformation projects by allowing for a fast and easy test of sgRNA accessibility in vivo ., (Copyright © 2020 Brandt, Gunn, Moretti and Zemetra.)

The following article asks about the speeches, practices, representations and experiences on health, hygiene and illnes in boarding schools early last century, taking for instance the School of Arts and Crafts of the Salesian Congregation, with the intention of revisiting the dialogue between medicine, hygiene and school in the context of development of the social question, the result of accelerated modernization process experienced by the city of Córdoba.

Purpose: Congenital hemophilia is a hereditary bleeding disorder that affects 1 in 5,000 males and is characterized by repetitive musculoskeletal bleeding episodes. Selective embolization of the knee and elbow arteries can prevent bleeding episodes. To evaluate the long-term efficacy of these procedures, we assessed the outcomes of 30 procedures performed in our center., Methods: We performed 30 procedures in 27 hemophilic patients, including 23 knee, and 7 elbow procedures. To evaluate the efficacy of selective embolization of knee and elbow arteries in people with hemophilia, we analyzed the number of bleeding episodes during 12 months before the procedure compared with the amount of episodes that occurred 3, 6, and 12 months after embolization., Results: Twenty-nine of 30 procedures were classified as successful. The median of 1.25 episodes per month (range 0-3) observed before the procedure was reduced to 0 (range 0-1.67; p < 0.001) at 3 months, 0.17 (range 0-1.67; p < 0.001) at 6 months, and 0.33 (range 0-1.67; p = 0.024) at 12 months. Three patients remained free of bleeding events for more than 6 months. Additionally, after the procedure there was a significant reduction in factor FVIII usage that sustained up to 12 months after the procedures. No serious adverse events were observed., Conclusions: Selective angiographic embolization of knee and elbow arteries is a feasible procedure that can prevent repetitive bleedings, which would translate in better joint outcomes for these patients.

A live chimeric vaccine virus against Japanese encephalitis (JE), ChimeriVax-JE, was used to define methods for optimal, random insertion of foreign immunologic determinants into flavivirus glycoproteins. The conserved M2e peptide of influenza A virus was randomly inserted into the yellow fever-specific NS1 glycoprotein of ChimeriVax-JE. A technique combining plaque purification with immunostaining yielded a recombinant virus that stably expressed M2e at NS1-236 site. The site was found permissive for other inserts. The insertion inhibited NS1 dimerization in vitro, which had no significant effect on virus replication in vitro and immunogenicity in vivo. Two different NS1-specific monoclonal antibodies and a polyclonal antibody efficiently recognized only the NS1 protein dimer, but not monomer. Adaptation of the virus to Vero cells resulted in two amino acid changes upstream from the insert which restored NS1 dimerization. Immunized mice developed high-titer M2e-specific antibodies predominantly of the IgG2A isotype indicative of a Th1-biased response.

An altered interaction between circulating LDL and endothelial cells might be at the basis of the increased prevalence of atherosclerosis in diabetes mellitus. The aim of the present work was to investigate the effect of a short incubation period with LDL from Type 1 diabetic patients in good metabolic control on endothelial cells derived from human aorta (HAEC). Cultured HAEC were incubated for 3 h with culture medium alone (control HAEC), with native LDL from healthy subjects (control LDL), or with native LDL from Type 1 diabetic patients (Type 1 LDL). After the incubation the following parameters were evaluated: endothelial cell nitric oxide synthase (NOS) activity, nitric oxide (NO) and peroxynitrite production, Na(+)/K(+)-ATPase and Ca(2+)-ATPase activities, intracellular Ca(2+) concentration and fluidity of the superficial part of the plasma membrane studied by 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH). Moreover, we studied the cellular activation, evaluated by the fluid phase endocytosis of TMA-DPH, and the microetherogeneity of the membrane surface, evaluated by dynamic fluorescence. HAEC incubated with control LDL showed compared with control HAEC: increased anisotropy and exponential lifetime of TMA-DPH, and enhanced TMA-DPH internalization. HAEC incubated with Type 1 LDL showed compared with both control HAEC and HAEC incubated with control LDL: (i) increased Na(+)/K(+)-ATPase and Ca(2+)-ATPase activities, and intracellular Ca(2+) concentration; (ii) increased NOS activity, NO and peroxynitrite production; (iii) increased anisotropy of TMA-DPH; (iv) enhanced internalization of the probe. The exponential lifetime and the width of distribution of TMA-DPH were significantly increased by Type 1 LDL only in comparison with control HAEC. The results suggest that a short-term interaction with LDL from Type 1 diabetic patients causes alterations of the plasma membrane surface and of cellular functions in endothelial cells in a possibly atherogenic way.

Aims/hypothesis: The molecular mechanisms involved in the platelet activation observed in hyperhomocysteinemia are not known. We aimed to discover if homocysteine concentrations are associated with abnormal platelet nitric oxide production in healthy and diabetic subjects., Methods: The study cohort included 28 patients with Type I (insulin-dependent) diabetes mellitus, 30 patients with Type II (non-insulin-dependent) diabetes mellitus, and 34 healthy subjects. Homocysteine plasma concentrations were measured by high-performance liquid chromatography. Platelet nitric oxide production was measured using a nitric oxide meter before and after a 3-h incubation with 100 micromol/l homocysteine. Stimulation experiments were done in vitro by the addition of alpha-thrombin (0.2 U/ml)., Results: Basal platelet nitric oxide production was lower in diabetic patients than in healthy subjects. Nitric oxide release was reduced by in vitro homocysteine incubation, being lower in platelets from diabetic patients than in platelets from control subjects. Thrombin increased nitric oxide synthesis in platelets from healthy subjects both in the presence and absence of homocysteine. In diabetic subjects thrombin increased nitric oxide release in the absence of homocysteine. But in the presence of homocysteine the response was reduced. An inverse relation was found between plasma homocysteine levels and basal platelet nitric oxide release in diabetic and healthy subjects., Conclusion/interpretation: Homocysteine could exert its atherogenic action in healthy and diabetic subjects partly by inhibiting platelet nitric oxide production with the subsequent increased platelet activation and aggregation.

Decreased levels of nitric oxide (NO) may contribute to impaired endothelium-dependent vasodilatation in essential hypertension. Moreover, in hypertension, major platelets aggregation and endothelial adhesion, and increased atherogenetic risks are also present. Nitric oxide produced by platelet NO synthase, which is similar to endothelial NO synthase, inhibits platelets aggregation by increasing cytoplasmic cyclic GMP levels and contributes in a major way to the antithrombogenic properties of endothelium. The aim of this study was to investigate platelet NO production and cytosolic Ca2+ levels in patients with essential hypertension and in healthy subjects. We studied NO production in 36 subjects (21 patients had essential hypertension and 15 subjects were normotensive); NO synthase activity was evaluated by measuring nitrite levels by the Griess reaction in the supernatant of sonicated platelets. Cytosolic Ca2+ levels were measured in intact platelets using the fluorescent probe Fura 2-AM. Nitric oxide levels in platelets were found higher in normotensive than in hypertensive patients (P < .0001). Nitric oxide levels in hypertensive women were significantly higher than in hypertensive men (P < .001). Hypertensive women and men had lower levels of nitrite than normotensive women and men (P < .001 and P < .002, respectively). Platelet cytosolic Ca2+ levels were higher in hypertensive patients than in normotensive subjects (P < .001). An inverse correlation was found between platelet cytosolic Ca2+ and NO levels (r = 0.74, P < .002). These data confirm the link between hypertension and altered platelets function and suggest a role for NO in cardiovascular events. Moreover, the higher levels of nitric oxide in child-bearing age women than in men further support the protective effect of estrogens on cardiovascular diseases.

In this study, serotonin (5-hydroxytryptamine; 5HT)-immunoreactive (5HT-IR) neuronal fibers were identified in the primary olfactory pathway of the sea lamprey. These neurons are likely part of a nonolfactory neural system that innervates the olfactory sac. Cell bodies with 5HT immunoreactivity predominated in the lamina propria of the rostral portion of the nasal cavity and were less prevalent adjacent to the olfactory epithelium. The 5HT-IR fibers were parallel to axons of the olfactory receptor neurons in the lamina propria of the olfactory mucosa and in the olfactory nerve. Serotonergic fibers crossed from the olfactory nerve into the olfactory bulb or branched in the caudal portion of the olfactory nerve and terminated at the junction of the olfactory nerve with the olfactory bulb. In the dorsal olfactory bulb, 5HT-IR fibers coursed along the layer of olfactory fibers. Throughout the layer with glomeruli and mitral cells, 5HT-IR fibers were seen along the border of glomerular units. Experimental lesion of the olfactory nerve was used to determine the origin of 5HT-IR fibers rostral to the olfactory bulb. The loss of these fibers and their reappearance during outgrowth of olfactory receptor neurons inferred that they emanate from the cell bodies in the olfactory sac. The results from this study suggest that axons of olfactory receptor neurons in larval lampreys receive modulation by 5HT from these neuronal fibers., (Copyright 2000 Wiley-Liss, Inc.)

The aim of the present work was to analyze the effect of LDL obtained from type 1 diabetic patients in good metabolic control on human umbilical vein endothelial cells (HUVECs) after a short incubation period to detect possible atherogenic modifications of endothelial properties. Cultured HUVECs were incubated for 3 h with culture medium alone (control HUVEC), with native LDL from 12 healthy men (control LDL), or with native LDL from 12 type 1 diabetic men (type 1 LDL) (100 pg/ml). After the incubation, the following parameters were evaluated: nitric oxide synthase (NOS) activity, cytoplasmic Ca2+ levels, Na+-K+-ATPase activity, plasma membrane fluidity determined by means of 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH), and plasma membrane conjugated diene (CD) content. The same experiments were repeated after bradykinin stimulation or in the presence of the antioxidant butylated hydroxytoluene (BHT), and nitric oxide (NO) production in intact HUVECs was also evaluated. HUVECs incubated with control LDL in comparison with control HUVECs showed a decreased fluidity of the membrane surface evaluated by TMA-DPH and a higher CD content. These alterations were prevented by the presence of BHT. HUVECs incubated with type 1 LDL in comparison with both control HUVECs and cells incubated with control LDL showed 1) increased NOS and Na+-K+-ATPase activity, cytoplasmic Ca2+ levels, and CD content, and 2) decreased fluidity of the membrane surface evaluated by TMA-DPH. These modifications were blunted--but not abolished--by the presence of BHT. After bradykinin stimulation either in the absence or in the presence of BHT, both cytoplasmic Ca2+ levels and NO production were increased in control HUVECs and in HUVECs incubated with control LDL, while a reduced response was observed in HUVECs incubated with type 1 LDL. The alterations observed in the endothelial function after the cell-LDL interaction might play a central role in the atherogenic process in diabetes.

The aim of the present study was to evaluate the action of plasma from insulin-dependent diabetic (IDDM) pregnant women on nitric oxide synthase (NOS) activity in cultured human umbilical vein endothelial cells (HUVECs). We also studied the effect of the plasma on cytosolic calcium and on Na+/K+-adenosine triphosphatase (ATPase) activity. Dynamic fluorescence studies of membrane fluidity were contemporarily performed to detect a direct effect of plasma on the endothelial cell membrane. We observed a significant increase in NOS activity, intracellular calcium, and Na+/K+-ATPase activity in cultured HUVECs exposed to IDDM plasma. Our dynamic fluorescence study showed a different microenvironmental organization of the cellular membrane after incubation with plasma from IDDM pregnant women, with a marked decrease in microheterogeneity as evaluated in terms of 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) lifetime distribution width. The present investigation suggests that plasma from IDDM pregnant women can cause a generalized disturbance in the function of endothelial cells cultured from healthy subjects. Such a modification might play a central role in the pathogenesis of the vascular complications of the disease.

The production of Pertussis Vaccine was reevaluated at the Instituto Nacional de Higiene "Rafael Rangel" in order to optimise it in terms of vaccine yield, potency, specific toxicity and efficiency (cost per doses). Four different processes, using two culture media (Cohen-Wheeler and Fermentación Glutamato Prolina-1) and two types of bioreactors (25 L Fermentador Caracas and a 450 L industrial fermentor) were compared. Runs were started from freeze-dried strains (134 or 509) and continued until the obtention of the maximal yield. It was found that the combination Fermentación Glutamato Prolina-1/industrial fermentor, shortened the process to 40 hours while consistently yielding a vaccine of higher potency (7.91 +/- 2.56 IU/human dose) and lower specific toxicity in a mice bioassay. In addition, the physical aspect of the preparation was rather homogeneous and free of dark aggregates. Most importantly, the biomass yield more than doubled those of the Fermentador Caracas using the two different media and that in the industrial fermentor with the Cohen-Wheeler medium. Therefore, the cost per doses was substantially decreased.