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BackgroundThe oral cavity is one of the most complex human body environments. Indeed, the continuous variation of this habitat conditions reflects the high dynamism of the resident microbial community. Two key actors in the oral diseases are the bacteria Streptococcus mutans and Fusobacterium nucleatum, both implicated in the formation of oral biofilms and consequently in the generation of common pathologies such as caries and various gingival and soft tissue inflammation diseases. In addition, F. nucleatum is also implicated in the halitosis phenomenon, thanks to its demonstrated ability to produce as second metabolite the hydrogen sulphide (H2S), one of the volatile sulphur compounds (VSCs) that, with methyl mercaptan (CH3SH) and the dimethyl sulphide (CH3SCH3)24, is produced by periodontopathic anaerobic bacteria and causes the awkward bad breath in halitosis patients.MethodsIn this study, the oral preparation Vea® Oris constituted only by vitamin E and capric/caprylic acid was evaluated as a potential treatment of caries and periodontal diseases; the effect of the product at different concentrations on the growth and the ability of both strains to form biofilm was investigated. Regarding to F. nucleatum also the influence of Vea® Oris on the production of H2S was evaluated. ResultsOur in vitro results suggested that the Vea® Oris treatment could considerably reduce the growth and biofilm formation of both S. mutans and F. nucleatum. For F. nucleatum an appreciable reduction of the H2S production can be also obtained. ConclusionsOverall, this study highlighted the potential of Vea® Oris as a more “natural” adjuvant to prevent the biofilm and plaque formation and to reduce the smelly odour of halitosis.

Made available in DSpace on 2022-04-29T08:30:53Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-04-01 National Health and Medical Research Council Objective: To describe fluid resuscitation practices in Brazilian intensive care units and to compare them with those of other countries participating in the Fluid-TRIPS. Methods: This was a prospective, international, cross-sectional, observational study in a convenience sample of intensive care units in 27 countries (including Brazil) using the Fluid-TRIPS database compiled in 2014. We described the patterns of fluid resuscitation use in Brazil compared with those in other countries and identified the factors associated with fluid choice. Results: On the study day, 3,214 patients in Brazil and 3,493 patients in other countries were included, of whom 16.1% and 26.8% (p < 0.001) received fluids, respectively. The main indication for fluid resuscitation was impaired perfusion and/or low cardiac output (Brazil: 71.7% versus other countries: 56.4%, p < 0.001). In Brazil, the percentage of patients receiving crystalloid solutions was higher (97.7% versus 76.8%, p < 0.001), and 0.9% sodium chloride was the most commonly used crystalloid (62.5% versus 27.1%, p < 0.001). The multivariable analysis suggested that the albumin levels were associated with the use of both crystalloids and colloids, whereas the type of fluid prescriber was associated with crystalloid use only. Conclusion: Our results suggest that crystalloids are more frequently used than colloids for fluid resuscitation in Brazil, and this discrepancy in frequencies is higher than that in other countries. Sodium chloride (0.9%) was the crystalloid most commonly prescribed. Serum albumin levels and the type of fluid prescriber were the factors associated with the choice of crystalloids or colloids for fluid resuscitation. Discipline of Anesthesiology Pain and Intensive Care Universidade Federal de São Paulo, Rua Napoleão de Barros, 737, 6ºandar Critical Care and Trauma Division George Institute for Global Health Hospital Sírio-Libanês Hospital das Clínicas Faculdade de Medicina Universidade de São Paulo Instituto de Pesquisa HCor - Hospital do Coração Instituto de Pesquisa Clínica Evandro Chagas Fundação Oswaldo Cruz Hospital de Clínicas Mário Lioni Hospital do Rim e Hipertensão Fundação Oswaldo Ramos Hospital Samaritano Hospital Paulistano Hospital São José Instituto D’Or de Ensino e Pesquisa Hospital Municipal São José Hospital Israelita Albert Einstein Hospital Santa Rita Hospital de Base Faculdade de Medicina de São Jose do Rio Preto Hospital Unimed de Belo Horizonte Hospital Santa Lúcia Hospital Evangélico de Sorocaba Hospital PIO XII Hospital São Camilo Pompéia Hospital Alvorada Brasília Hospital Ecoville Hospital do Trabalhador Hospital Vita Batel Hospital das Clínicas Faculdade de Medicina de Botucatu Universidade Estadual “Júlio de Mesquita Filho” Hospital Geral Dr. César Cals Hospital Copa D’Or Associação Beneficente Hospital Unimar Hospital Casa de Saúde de Santos Hospital Estadual Getúlio Vargas Hospital e Maternidade Otaviano Neves Instituto Estadual do Cérebro Paulo Niemeyer Hospital Universitário Universidade Federal de Juiz de Fora Lifecenter Hospital Geral de Fortaleza Hospital Municipal Dr. Moysés Deutsch (M’Boi Mirim) Santa Casa de Misericórdia de Presidente Prudente Hospital 9 de julho Hospital Estadual Rocha Faria Hospital Municipal Santa Isabel Hospital e Maternidade Municipal Dr. Odelmo Leão Carneiro Hospital Evangélico de Londrina Universidade Estadual de Londrina Hospital Samaritano João Pessoa Hospital de Caridade Astrogildo de Azevedo Vitória Apart Hospital Instituto de Ortopedia e Traumatologia Hospital do Subúrbio Hospital Renascentista Hospital Felício Rocho Hospital Municipal Irmã Dulce Hospital Universitário Júlio Müller Universidade Federal do Mato Grosso Hospital de Urgência Hospital Geral de Goiânia Fundação Doutor Amaral Carvalho Hospital Adventista de Belém Hospital Santa Juliana Hospital Dom Vicente Scherer Hospital Primavera Hospital Carlos da Silva Lacaz Hospital Escola Faculdade de Medicina de Jundiaí Hospital Sepaco Hospital Professor Edmundo Vasconcelos Hospital Paulo Sacramento Clínica Dom Rodrigo Complexo Hospitalar Ortotrauma de Mangabeira Hospital Nossa Senhora da Conceição Hospital Regional de Samambaia Hospital São Camilo Ipiranga Hospital da Restauração Instituto do Coração Hospital das Clínicas Faculdade de Medicina Universidade de São Paulo Hospital Estadual Jayme Santos Neves Hospital SOS Cárdio Hospital da Luz Vila Mariana Hospital Maternidade e Pronto-Socorro Santa Luci Santa Casa de Misericórdia de Vitória da Conquista Santa Casa de Misericórdia de Porto Alegre Hospital São Francisco de Assis Hospital Memorial São José Hospital Regional de Jundiaí Hospital Universitário Faculdade de Medicina de Jundiaí Hospital Regional de Presidente Prudente Santa Casa de Misericórdia de Assis Hospital de Clínicas Gaspar Vianna Hospital do Coração do Brasil Hospital Hélio Anjos Ortiz Hospital Vila da Serra Hospital Nereu Ramos Hospital Santa Maria Intensibarra Santa Casa de Misericórdia de Santo Amaro Santa Casa de Caridade de Don Pedrito Santa Casa de Misericórdia de Santana do Livramento Hospital Unimed de Macaé Hospital Municipal Pedro II Hospital Federal dos Servidores do Estado Hospital São Mateus Hospital IBR Hospital Uniclinic Santa Casa de Misericórdia de Paraguaçu Paulista Associação Hospitalar Beneficente São Vicente de Paulo Hospital do Servidor Público Municipal de São Paulo Hospital Santa Isabel Hospital Municipal Dr Jose Soares Hungria Hospital Fernandes Távora Hospital Distrital Evandro Ayres de Moura Hospital Saúde da Mulher Hospital Estadual de Urgência e Emergência de Vitória Samur Hospital e Pronto-Socorro 28 de Agosto Hospital Assunção Hospital Universitário de Santa Maria Hospital Universitário Cajuru Instituto do Câncer do Estado de São Paulo Hospital Unimed de Limeira Hospital Amecor Santa Casa de Caridade de Diamantina Hospital das Clínicas Faculdade Ribeirão Preto Universidade de São Paulo HCor - Hospital do Coração Hospital Goiânia Leste Hospital Ortopédico Hospital Santa Maria Hospital Municipal Dr. Munir Rafful Hospital Jardim Amália Hospital Madre Regina Prottman Hospital Universitário São Francisco de Paula Universidade Católica de Pelotas Hospital São Joao de Deus Hospital Nossa Senhora Monte Serrat Hospital Unimed Salto Hospital Moinhos de Vento Hospital Geral de Vitória da Conquista Hospital Marcelino Champagnat Hospital São Lucas Pontifícia Universidade Católica do Rio Grande do Sul Hospital Universitário Universidade Federal da Grande Dourados Hospital Português Hospital Brigadeiro Hospital Regional de Sousa Hospital das Clínicas Universidade Federal do Espírito Santo Santa Casa de Misericórdia de Ribeirão Preto Hospital Universitário Lauro Wanderley Santa Casa de Belo Horizonte Hospital Adventista de Manaus Santa Casa Maringá Universidade Estadual Maringá Hospital Total Cor Hospital Universitário Universidade de Santa Cruz do Sul Hospital Dom Hélder Hospital das Clínicas de Porto Alegre Universidade Federal do Rio Grande do Sul Hospital Anis Rassi Instituto de Infectologia Emílio Ribas II Hospital Unimed Rio de Janeiro Hospital Alemão Oswaldo Cruz Hospital Samaritano Rio de Janeiro São Bernardo Apart Hospital Hospital Nossa Senhora dos Prazeres Hospital Unimed ABC Hospital Municipal de Paracatu Hospital Municipal Padre Germano Lauck Hospital Santa Helena Hospital Santa Izabel Santa Casa de Misericórdia de São Paulo Hospital Tereza Ramos Hospital Alvorada Taguatinga Hospital Maternidade São José Hospital Universitário Ciências Médicas Fundação Educacional Lucas Machado Santa Casa de Belém do Pará Instituto Nacional de Cardiologia Hospital Novo Atibaia BP - A Beneficência Portuguesa de São Paulo Hospital Universitário Maria Aparecida Pedrossian Universidade Federal de Mato Grosso do Sul Hospital do Servidor Público Estadual “Francisco Morato de Oliveira” Hospital Norte D’Or Hospital Estadual Ipiranga Hospital Universitário de Maringá Universidade Estadual de Maringá Albert Sabin Hospital e Maternidade Casa de Caridade de Carangola Irmandade de Misericórdia de Guaxupé Disciplina de Emergências Clínicas Universidade Estadual de Campinas Hospital São Lucas Fundação Pio XII- Hospital de Câncer de Barretos Clínica Campo Grande Hospital das Clínicas Universidade Federal de Minas Gerais Casa de Saúde Santa Lúcia Hospital Regional de Itapetininga São Camilo Santa Casa de Angra dos Reis Grupo Hospitalar Nossa Senhora da Conceição Irmandade Misericórdia Hospital Santa Casa de Monte Alto Hospital São Marcos Hospital Unimed de Manaus Hospital Universitário Getúlio Vargas Universidade Federal do Amazonas Casa de Saúde Campinas Hospital e Maternidade Galileo Hospital das Clínicas Faculdade de Medicina de Botucatu Universidade Estadual “Júlio de Mesquita Filho” National Health and Medical Research Council: APP1039312

PURPOSE: The miniaturized transcatheter pacing system (TPS) implant is performed using a 27 Fr sheath. Achieving femoral vein access hemostasis after sheath removal is of utmost importance. Feasibility and clinical effectiveness of double device-based suture-mediated closure technique (DualPerclose) were evaluated. METHODS: Patients undergoing TPS positioning and treated with DualPerclose technique at our institution were considered. Feasibility of the DualPerclose technique included the rates of effective initial device suture fixation and effective hemostasis after sheath removal. Clinical efficacy considered intraprocedural and periprocedural bleeding events as well as midterm access site vascular injury assessed at 3 months using lower limb vascular ultrasonography. Data on all follow-up major adverse events were also collected. RESULTS: All patients (n = 83 patients; mean age 82.3 ± 7.1 years, 67.5% male gender, 85.5%, with structural heart disease, mean left ventricular ejection fraction 54.0 ± 9.6%, renal impairment in 31.3%) who underwent TPS positioning between November 2015 and February 2020 were considered. TPS positioning was successful in all patients. In 82 patients, the DualPerclose approach was utilized: 13 patients (15.8%) required > 2 devices to obtain effective fixing of 2 sutures; complete immediate hemostasis was achieved in 80 patients (97.6%). One (1.2%) severe groin-related bleeding event occurred. At midterm, 1 (1.2%) mildly symptomatic arteriovenous fistula was diagnosed (conservative treatment). Over a median follow-up of 22 (IQR 10-35.5) months, 14 (17.1%) major adverse events were recorded, including 1 loss of capture requiring TPS replacement and 5 deaths. CONCLUSION: Femoral vein access closure using dual Perclose Proglide devices during TPS is feasible and clinically effective.

During the Coronavirus Disease 2019 worldwide pandemic, patients with heart failure are a high-risk group with potential higher mortality if infected. Although lockdown represents a solution to prevent viral spreading, it endangers regular follow-up visits and precludes direct medical assessment in order to detect heart failure progression and optimize treatment. Furthermore, lifestyle changes during quarantine may trigger heart failure decompensations. During the pandemic, a paradoxical reduction of heart failure hospitalization rates was observed, supposedly caused by patient reluctance to visit emergency departments and hospitals. This may result in an increased patient mortality and/or in more complicated heart failure admissions in the future. In this scenario, different telemedicine strategies can be implemented to ensure continuity of care to patients with heart failure. Patients at home can be monitored through dedicated apps, telephone calls, or devices. Virtual visits and forward triage screen the patients with signs or symptoms of decompensated heart failure. In-hospital care may benefit from remote communication platforms. After discharge, patients may undergo remote follow-up or telerehabilitation to prevent early readmissions. This review provides a comprehensive appraisal of the many possible applications of telemedicine for patients with heart failure during Coronavirus disease 2019 and elucidates practical limitations and challenges regarding specific telemedicine modalities. JD is a researcher for the Limburg Clinical Research Center (LCRC) UHasselt-ZOL-Jessa, supported by the foundation Limburg Sterk Merk (LSM), province of Limburg, Flemish government, Hasselt University, Ziekenhuis Oost-Limburg, and Jessa Hospital. Tersalvi, G (corresponding author), Fdn Cardioctr Ticino, Div Cardiol, Lugano, Switzerland. Hirslanden Klin St Anna, Dept Internal Med, Luzern, Switzerland. tersalvi@gmail.com

International audience; In 2005 the then ESA Directorate for Human Spaceflight, Microgravity and Exploration (D-HME) commissioned a study from the European Science Foundation's (ESF) European Space Sciences Committee (ESSC) to examine the science aspects of the Aurora Programme in preparation for the December 2005 Ministerial Conference of ESA Member States, held in Berlin. A first interim report was presented to ESA at the second stakeholders meeting on 30 and 31 May 2005. A second draft report was made available at the time of the final science stakeholders meeting on 16 September 2005 in order for ESA to use its recommendations to prepare the Executive proposal to the Ministerial Conference. The final ESSC report on that activity came a few months after the Ministerial Conference (June 2006) and attempted to capture some elements of the new situation after Berlin, and in the context of the reduction in NASA's budget that was taking place at that time; e. g., the postponement sine die of the Mars Sample Return mission. At the time of this study, ESSC made it clear to ESA that the timeline imposed prior to the Berlin Conference had not allowed for a proper consultation of the relevant science community and that this should be corrected in the near future. In response to that recommendation, ESSC was asked again in the summer of 2006 to initiate a broad consultation to define a science-driven scenario for the Aurora Programme. This exercise ran between October 2006 and May 2007. ESA provided the funding for staff support, publication costs, and costs related to meetings of a Steering Group, two meetings of a larger ad hoc group ( 7 and 8 December 2006 and 8 February 2007), and a final scientific workshop on 15 and 16 May 2007 in Athens. As a result of these meetings a draft report was produced and examined by the Ad Hoc Group. Following their endorsement of the report and its approval by the plenary meeting of the ESSC, the draft report was externally refereed, as is now normal practice with all ESSC-ESF reports, and amended accordingly. The Ad Hoc Group defined overarching scientific goals for Europe's exploration programme, dubbed "Emergence and co-evolution of life with its planetary environments,"focusing on those targets that can ultimately be reached by humans, i.e., Mars, the Moon, and Near Earth Objects. Mars was further recognized as the focus of that programme, with Mars sample return as the recognized primary goal; furthermore the report clearly states that Europe should position itself as a major actor in defining and leading Mars sample return missions. The report is reproduced in this article. On 26 November 2008 the Ministers of ESA Member States decided to give a high strategic priority to the robotic exploration programme of Mars by funding the enhanced ExoMars mission component, in line therefore with the recommendations from this ESSC-ESF report.

We present a survey of the introduction and evolution of microscopy techniques in Spain, and the concepts and lines of research developed around this instrument, particularly in the field of Biomedical research. We cover in our article the long period from the XVII Century to the arrival of the great figure of Santiago Ramon y Cajal (1853-1934). We particularly want to mention many of the previously neglected pioneers who certainly paved the route for his discoveries and, we believe that without them, he would never have arrived to his important position in the annals of Biology and Medicine. The historical, scientific and social framework of that period also helped the approach to important biological concepts such as the cell and tissue, which are previous and essential ideas for a correct understanding of Development.

6 páginas, 2 figuras, 1 tabla -- PAGS nros. 299-304, In the GENE experiment performed during an 11-day Soyuz Mission to the International Space Station (ISS), we intended to determine if microgravity affects Drosophila metamorphosis processes. Control experiments were performed including a 1g ground control parallel to the ISS flight samples and a Random Position Machine microgravity simulated control. A preliminary analysis of the results indicates that five hundred to one thousand genes change their expression profiles depending on the cut-off levels selected. Especially affected among them are the mitochondrial ones (an example with the respiratory chain is presented). We show here that there is a synergic effect of the constraints introduced to meet the requirements of the space experiment (mainly, a cold step and the use of hermetically closed Type-I containers). The cold transport step to the launch site was introduced to slow down the pupal development. The hermetically closed Type I containers were required to ensure the containment of the fixative (acetone) in the experiment. As shown here, the oxygen concentration inside the container was not optimal but fully compatible with pupal development. It is highly likely that such combined environmental effects will become a common finding in these types of studies as they become more complicated and extensive. They could open the way to understand how the gene expression patterns and the actual phenotypes can adjust to the environment. These findings indicate the importance of a vigorous ground based program in support of real microgravity experiments. Only then we can utilize the ISS in order to understand the consequences of the modified environment in outer space on living organisms, This work was supported by Grants from The Spanish "Plan Nacional de Investigación Científica y Desarrollo Tecnológico" Ref. nos. ESP2001-4522-PE, and ESP2003-09475-C02-01 and from The Netherlands Institute for Space Researchs, NW0-SROM, MG-057

5 páginas -- PAGS nros. 293-297, Changes have been reported in the pattern of gene expression in Arabidopsis on exposure to microgravity. Plant cell growth and proliferation are functions that are potentially affected by such changes in gene expression. In the present investigation, the cell proliferation rate, the regulation of cell cycle progression and the rate of ribosome biogenesis (this latter taken to estimate cell growth) have been studied using morphometric markers or parameters evaluated by light and electron microscopy in real microgravity on the International Space Station (ISS) and in ground-based simulated microgravity, using the Random Positioning Machine and the Magnetic Levitation Instrument. Results showed enhanced cell proliferation but depleted cell growth in both real and simulated microgravity, indicating that the two processes are uncoupled, unlike the situation under normal gravity on Earth in which they are strictly co-ordinated events. It is concluded that microgravity is an important stress condition for plant cells compared to normal ground gravity conditions

Neurobiological experiments on 4 animal species (Xenopus laevis, Pleurodeles waltl, Drosophila melanogaster, Acheta domesticus) were performed to study effects of microgravity on development and aging of neuronal, sensory and motor systems. Animal models were selected according to their suitability to answer questions concerning μg-effects on neuroanatomy, neuronal activity, and behaviour. The studies were performed on the Soyuz Taxi flights Andromede, Cervantes, Eneide and LDM-TMA8/TMA7. Observations from these flights include: (1) In tadpoles and cricket larvae, morphological features of sensory cells and neurons are rarely affected by microgravity. (2) In crickets, in-flight fertilization was successful; after landing, flight larvae hatched earlier than ground reared siblings. (3) In crickets, proliferation of peptidergic neurons and their projection patterns within the nervous system were not affected by microgravity. (4) During aging, the impact of microgravity on peptidergic neurons of male Drosophila was limited to the size of cell body. (5) In Xenopus, neurophysiological features of the spinal motor system during fictive swimming were partially modified. (6) In Xenopus tadpoles, the vestibuloocular reflex was affected in an age-related manner. Modifications were also related to the occurrence of a tail lordosis induced by microgravity. It is concluded that adaptation to microgravity during development and aging is mainly based on physiological mechanisms within the central nervous system while structural modifications of the sensory and neuronal system contribute less.

In an experiment conducted in the “Cervantes” Spanish Soyuz Mission, a 10-day flight to the International Space Station, Arabidopsis seeds were germinated, seedlings grew for 4 days at 22ºC, and they were fixed in flight with paraformaldehyde. A ground 1 g control experiment was replicated, and an additional experiment in simulated microgravity, using a Random Positioning Machine, was performed in the same conditions. Structural, morphometric and immunocytochemical data were compared. Glutaraldehyde-fixed seedlings similarly grown in the Space Shuttle (STS-84 Mission) were also used for ultrastructural and morphometric studies. Seedlings grown for 4 days in real or simulated weightlessness showed a longer size than the ground 1 g control. Root meristematic cells showed an enhanced proliferating rate, but ribosome biogenesis was reduced, as inferred from the nucleolar size and from the levels of the nucleolar protein nucleolin. This could be the consequence of the acceleration of the cell cycle, with shortening of its phases. Weightlessness induces stress in the plant, influencing cellular processes decisive for development and morphogenesis. This stress may endanger the plant and would require the action of compensating specialized defence mechanisms

Aim Transthoracic echocardiography (TTE) during cardiopulmonary arrest (CPA) has been studied in victims of cardiac arrests. Our objective was to evaluate the feasibility and usefulness of TTE in victims of cardiac arrest with non-shockable rhythms hospitalized in intensive care units (ICUs). Methods This prospective and observational cohort study evaluated ICU patients with CPA in asystole or pulseless electrical activity (PEA). Intensivists performed TTE during intervals of up to 10 s as established in the treatment protocol. Myocardial contractility was defined as intrinsic movement of the myocardium coordinated with cardiac valve movement. PEA without contractility was classified as electromechanical dissociation (EMD), and with contractility as pseudo-EMD. The images, the rates of return of spontaneous circulation (ROSC) and the survival upon hospital discharge and after 180 days were evaluated. Results A total of 49 patients were included. Image quality was considered adequate in all cases and contributed to the diagnosis of CPA in 51.0% of the patients. Of the 49 patients included, 17 (34.7%) were in asystole and 32 (65.3%) in PEA, among which 5 (10.2%) were in EMD and 27 (55.1%) in pseudo-EMD. The rates of ROSC were 70.4% for those in pseudo-EMD, 20.0% for those in EMD, and 23.5% for those in asystole. Survival upon hospital discharge and after 180 days occurred only in patients in pseudo-EMD (22.2% and 14.8%, respectively). Conclusions TTE conducted during cardiopulmonary resuscitation in ICU patients can be performed without interfering with care protocols and can contribute to the differential diagnosis of CPA and to the identification of a subgroup of patients with better prognosis.

In Drosophila melanogaster two high molecular weight tropomyosin isoforms, historically named heavy troponins (TnH-33 and TnH-34), are encoded by the Tm1 tropomyosin gene. They are specifically expressed in the indirect flight muscles (IFM). Their N-termini are conventional and complete tropomyosin sequences, but their C-termini consist of different IFM-specific domains that are rich in proline, alanine, glycine and glutamate. The evidence indicates that in Diptera these IFM-specific isoforms are conserved and are not troponins, but heavy tropomyosins (TmH). We report here that they are post-translationally modified by several phosphorylations in their C-termini in mature flies, but not in recently emerged flies that are incapable of flight. From stoichiometric measurements of thin filament proteins and interactions of the TmH isoforms with the standard Drosophila IFM tropomyosin isoform (protein 129), we propose that the TmH N-termini are integrated into the thin filament structural unit as tropomyosin dimers. The phosphorylated C-termini remain unlocated and may be important in IFM stretch-activation. Comparison of the Tm1 and Tm2 gene sequences shows a complete conservation of gene organisation in other Drosophilidae, such as Drosophila pseudoobscura, while in Anopheles gambiae only one exon encodes a single C-terminal domain, though overall gene organization is maintained. Interestingly, in Apis mellifera (hymenopteran), while most of the Tm1 and Tm2 gene features are conserved, the gene lacks any C-terminal exons. Instead these sequences are found at the 3' end of the troponin I gene. In this insect order, as in Lethocerus (hemipteran), the original designation of troponin H (TnH) should be retained. We discuss whether the insertion of the IFM-specific pro-ala-gly-glu-rich domain into the tropomyosin or troponin I genes in different insect orders may be related to proposals that the IFM stretch activation mechanism has evolved independently several times in higher insects., This research was supported by grants from the Spanish Government PB96-0069, ESP1999-0379-C02, ESP-2001-4521-PE, ESP2002-11913-E and ESP2003-09475-C02-01.

The proteolytic activites of Artemia salina immediately after hatching are found masked in a precursor of high molecular weight (∼ 100000). The molecular weight of this precursor decreases slightly as development proceeds. However, its kinetic and chromatographic properties vary greatly. Shortly after hatching, the activities are highly inhibited, can be activated severalfold by the chaotropic agent potassium iodide and are not proportional to the amount of enzyme-containing extract added. Later after hatching, its kinetics become normal. From these observations we have concluded that (a) the majority, if not all, of the proteinase activities found later in development are already present immediately after hatching, although in an inhibited state that is gradually activated; (b) the proteinases are not found free in the cytoplasm but in a complex which may allow regulation of their activity., This work has been supported by grants of the J. March Foundation and Fondo Nacional para la Investigación Científica R. G. is a fellow of the Consejo Superior de Investigaciones Científicas.

We explored the use of microwave technology in fixation with the objective of achieving quicker fixation regimes, lower concentrations of toxic and volatile reagents, and enhanced antigen detection. We used a modified domestic microwave oven (900 W) and a low-power (5 W) microwave bench. The work was done on plant materials. The oven was supplemented with a cooling device, a stirring system, and a record of the sample temperature and the time of effective irradiation. The sample, immersed in a fixative solution of 1% paraformaldehyde (PFA) in PBS, was irradiated for only 10 minutes. The sample temperature did not exceed 37 degrees C. In these mild conditions, the quality of the (ultra)structural preservation of the samples, morphometrically assessed, was at the same level as obtained with the same fixative, using conventional methods. On the contrary, samples fixed in the same conditions without irradiation showed a poor structural preservation. The antigenic preservation of the irradiated samples was excellent, since the labeling levels of two nucleolar proteins, detected by immunogold, were three times higher than in conventionally fixed samples. In the so-called microwave bench, the pathway of microwaves is guided, so that low-power microwaves directly hit the sample and there is no dispersion of energy. Temperature of fixative did not increase after microwave irradiation. Fixation in the bench with either 4% PFA, or 1% PFA, for 20 minutes resulted in structural preservation of samples similar in quality as obtained with conventional fixation and in a similar or better level of antigen preservation. Therefore, controlling temperature and effective irradiation is crucial in order to obtain optimal structural and antigen preservation with microwave-enhanced fixation. The dramatic differences observed between microwave-irradiated samples and samples fixed in the same conditions without irradiation, strongly support the existence of specific effects of microwaves on fixation, independent from the mere heating of the samples.

Few data are available on enzyme activity in amphibian plasma or erythrocytes. We measured the activity of several blood enzymes in the urodele amphibian Pleurodeles waltl reared under standard laboratory conditions. In subsequent experiments, we will estimate and compare the physiological and biochemical conditions of P. waltl when reared under extreme temperature or microgravity conditions. The enzymes selected were glutamate dehydrogenase, aspartate aminotransferase, alanine aminotransferase, superoxide dismutase, catalase, isocitrate dehydrogenase and glucose-6-phosphate dehydrogenase. In fresh plasma samples, enzyme activity in females was higher than in males, except for aspartate and alanine aminotransferases, which were equivalent in females and males. Glutamate dehydrogenase activity was higher in males than in females. In female erythrocytes, the activity of all enzymes was higher than in male erythrocytes. We have also studied the storage conditions of samples and observed that for most enzymes, the activity in freshly isolated plasma and erythrocyte preparations decreased after storage at −18 or +4°C.

To define the transcriptional mechanisms contributing to stage- and tissue-specific expression of muscle genes, we performed transgenic analysis of Drosophila paramyosin gene regulation. This gene has two promoters, one for paramyosin and one for miniparamyosin, which are active in partially overlapping domains. Regions between -0.9 and -1.7 kilobases upstream of each initiation site contribute to the temporal and spatial expression patterns. By comparing the Drosophila melanogaster and Drosophila virilis promoters, conserved binding sites were found for known myogenic factors, including one MEF2 site and three E boxes. In contrast with previous data, our experiments with the paramyosin promoter indicate that the MEF2 site is essential but not sufficient for proper paramyosin gene transcription. Mutations in the three E boxes, on the other hand, do not produce any effect in embryonic/larval muscles. Thus MEF2 site- and E box-binding proteins can play different roles in the regulation of different muscle-specific genes. For the miniparamyosin promoters, several conserved sequences were shown to correspond to functionally important regions. Our data further show that the two promoters work independently. Even when both promoters are active in the same muscle fiber, the transcription driven by one of the promoters is not affected by transcription driven by the other.

The complete genomic organization of the Drosophila troponin T (TnT) gene shows many interesting features, including the presence of a microexon of only 3 nucleotides conserved among Drosophilidae. It is the smallestbona fide exon so far described, placing a new lower limit on the nucleotide number required for correct splicing. Four muscle-type specific transcripts are generated by developmentally regulated alternative splicing. Exons 3, 4, and 5 are absent in the transcript present in jump and flight muscles. A total of 11 exons are present in the adult hypodermic muscles transcript, whereas the microexon is absent in the larval hypodermic musculature. The two isoforms differ in a lysine residue. Post-translational regulation of the flight muscles/tergal depressor of the trochanter-specific isoform is involved in flight and/or jump function. The interaction domains of TnT in the tropomyosin-troponin complex are strongly conserved in the known vertebrate and invertebrate TnT sequences, whereas the terminal regions show an important variability. The COOH-terminal region shows important phylogenetic variations, whereas the NH2-terminal domain is associated with specific muscle types in a particular organism, a finding that discloses a selective value for these domains in the functionality of distinct muscles in different organisms.

Two sets of muscle polypeptides showing calcium-binding capacity and intense labelling in vivo with 32P were purified and characterized from Drosophila melanogaster adult extracts. The polypeptides exhibit crossed immunoreactivity and share similar biochemical properties such as those involved in purification. They have been identified as isoforms of troponin-T (TnT) by sequence analysis of a cDNA clone isolated from an embryonic library. The two sets of TnT polypeptides correspond to the fibrillar and non-fibrillar muscle isoforms, respectively. The non-fibrillar muscle isoforms separate into two bands which are differentially expressed during development. Analysis of TnT isoforms in bee thoraces indicates that the expression of the fibrillar muscle isoform correlates with the acquisition of functional flight capability. In vivo labelling experiments reveal that the two TnT sets are readily phosphorylated. The Drosophila TnTs show calcium-binding properties by three different types of assays. Our results suggest that this property could be specific to insect TnTs and may be related to the long, extremely acidic polyglutamic carboxy-terminus present in these polypeptides, which does not occur in non-arthropod TnTs.

Standard viable preservation methods for biological samples using low temperatures have been investigated concerning their storage capabilities under higher temperature levels than usual. For a representative set of organism classes (plants, mammalian cells, arthropods and aquatic invertebrates), the minimum appropriate storage conditions have been identified by screening storage temperatures at -196 degrees, -80 degrees, -20 degrees, +4 degrees, +20 degrees/25 degrees C for periods from 2 days to 4 weeks. For storage below 0 degree C, as a typical cryopreservative, dimethylsulfoxide (DMSO) was used. For some samples, the addition of trehalose (as cryopreservative) and the use of a nitrogen atmosphere were investigated. After storage, the material was tested for vitality. The findings demonstrated that acceptable preservation can be achieved under higher storage temperatures than are typically applied. Small, dense cultured plant cells survive for 21 d when moderately cooled (+4 degrees to -20 degrees C); addition of trehalose enhances viability at -20 degrees C. For mammalian cells, the results show that human lymphocytes can be preserved for 3 d at 25 degrees C, 7 d at 4 degrees C and 28 d at -80 degrees C. Friend leukaemia virus transformed cells can be stored for 3 d at 25 degrees C, 14 d at 4 degrees C and 28 d at -80 degrees C. Hybridoma cells can be kept 7 d at 4 degrees C and 28 d at -20 degrees C or -80 degrees C. Model arthropod systems are well preserved for 2 weeks if maintained at lower temperatures that vary depending on the species and/or stage of development; e.g., 12 degrees C for Drosophila imagoes and 4-6 degrees C for Artemia nauplii. For aquatic invertebrates such as sea urchins, embryonic and larval stages can be preserved for several weeks at +6 degrees C, whereas sperm and eggs can best be stored at + 4 degrees C for up to 5 d at maximum. These results enhance the range of feasible space experiments with biological systems. Moreover, for typical terrestrial preservation methods, considerable modification potential is identified.