Matkapuhelinverkon MAP-protokollatoteutus älyverkkopalveluiden ohjauspisteeseen

Työssä esitellään yleiseurooppalaisen GSM-matkapuhelinjärjestelmän verkkoelementtejä ja perehdytään niiden väliseen standardoituun merkinantoprotokollaan. Lisäksi tarkastellaan protokollan lyhytsanomien välitykseen liittyviä operaatioita ja niissä tapahtunutta kehitystä standardoinnin eri vaiheissa. Tavoitteena oli toteuttaa GSM-matkapuhelinverkon merkinantoprotokollaan perustuva ohjelma, jonka tehtävänä on välittää lyhytsanomia matkapuhelin- ja lyhytsanomakeskuksen välillä. Matkapuhelimeen päättyvän lyhytsanoman välitykseen liittyy lisäksi reititystiedon hakeminen vastaanottajan kotirekisteristä. Toteutuksessa on ohjelmointirajapinta, joka helpottaa matkapuhelinverkon uusien palvelusovellusten kehittämistä. Toteutus testattiin standardoituja testitapauksia soveltaen. Yhdenmukaisuustestauksessa käytettiin apuna merkinantoanalysaattoria. Testauksessa tarkastettiin, että protokolla toimii loogisesti oikein. Suorituskykyä ei ole voitu testata todellisessa testiympäristössä, mutta ohjelmallisesti toteutettujen simulaattoreiden avulla on saatu hyviä tuloksia.

Comparison of central macular thickness (CMT) measurement between time domain and spectral domain (SD) optical coherence tomography (OCT) and reproducibility of SD OCT in active neovascular AMD

Purpose: To evaluate the reproducibility of Cirrus-SD OCT measurements and to compare central macular thickness (CMT) measurements between TD-Stratus and SD-Cirrus OCT in patients with active exudative AMD. Methods: Consecutive case series of patients with active exudative AMD seen in the Medical Retina Department. Patients underwent 1 scan with Stratus (macular thickness map protocol) and 5 scans with Cirrus (Macular Cube protocol) at the same visit by the same experienced examiner. To be included, patients best-corrected visual acuity (BCVA) had to be >20/200 while all scans had to be of sufficient quality, well-centered and at least one Cirrus scan with CMT >300 microns. The repeatability of the SD Cirrus was estimated by using all 5 CMT measurements and the mean of the Cirrus measurements was compared with the CMT obtained by TD Stratus. Results: Cirrus OCT demonstrated high intraobserver repeatability at the central foveal region (ICC 96%). The mean of the CMT measurements was 321microns for Stratus and 387 microns for Cirrus. The average difference was 65m (SD=30). The coefficient of concordance between Stratus and Cirrus CMT measurements was rho=0,749 with a high precision and a moderate accuracy. The equation of the line of regression between Stratus and meanCirrus is given by the following: M_stratus = 0,848 x m_cirrus - 4,496 (1).Conclusions: The Cirrus macular cube protocol allows reproducible CMT measurements in patients with active exudative AMD. In cases of upgrading from TD to SD use and vice versa, there is the possibility to predict the measurements by using the equation (1). These real life data and conclusions can help in improving our clinical management of patients with neovascular AMD.

Application of dissociation curve analysis to radiation hybrid panel marker scoring: generation of a map of river buffalo (B-bubalis) chromosome 20

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Probability sampling protocol of classification maps from spaceborne/airborne image

To deliver sample estimates provided with the necessary probability foundation to permit generalization from the sample data subset to the whole target population being sampled, probability sampling strategies are required to satisfy three necessary not sufficient conditions: (i) All inclusion probabilities be greater than zero in the target population to be sampled. If some sampling units have an inclusion probability of zero, then a map accuracy assessment does not represent the entire target region depicted in the map to be assessed. (ii) The inclusion probabilities must be: (a) knowable for nonsampled units and (b) known for those units selected in the sample: since the inclusion probability determines the weight attached to each sampling unit in the accuracy estimation formulas, if the inclusion probabilities are unknown, so are the estimation weights. This original work presents a novel (to the best of these authors' knowledge, the first) probability sampling protocol for quality assessment and comparison of thematic maps generated from spaceborne/airborne Very High Resolution (VHR) images, where: (I) an original Categorical Variable Pair Similarity Index (CVPSI, proposed in two different formulations) is estimated as a fuzzy degree of match between a reference and a test semantic vocabulary, which may not coincide, and (II) both symbolic pixel-based thematic quality indicators (TQIs) and sub-symbolic object-based spatial quality indicators (SQIs) are estimated with a degree of uncertainty in measurement in compliance with the well-known Quality Assurance Framework for Earth Observation (QA4EO) guidelines. Like a decision-tree, any protocol (guidelines for best practice) comprises a set of rules, equivalent to structural knowledge, and an order of presentation of the rule set, known as procedural knowledge. The combination of these two levels of knowledge makes an original protocol worth more than the sum of its parts. The several degrees of novelty of the proposed probability sampling protocol are highlighted in this paper, at the levels of understanding of both structural and procedural knowledge, in comparison with related multi-disciplinary works selected from the existing literature. In the experimental session the proposed protocol is tested for accuracy validation of preliminary classification maps automatically generated by the Satellite Image Automatic MapperT (SIAMT) software product from two WorldView-2 images and one QuickBird-2 image provided by DigitalGlobe for testing purposes. In these experiments, collected TQIs and SQIs are statistically valid, statistically significant, consistent across maps and in agreement with theoretical expectations, visual (qualitative) evidence and quantitative quality indexes of operativeness (OQIs) claimed for SIAMT by related papers. As a subsidiary conclusion, the statistically consistent and statistically significant accuracy validation of the SIAMT pre-classification maps proposed in this contribution, together with OQIs claimed for SIAMT by related works, make the operational (automatic, accurate, near real-time, robust, scalable) SIAMT software product eligible for opening up new inter-disciplinary research and market opportunities in accordance with the visionary goal of the Global Earth Observation System of Systems (GEOSS) initiative and the QA4EO international guidelines.

A comparison of landslide inventories along the shoreline of Steamboat Island Peninsula, Thurston County, WA: office based protocol vs. field based protocol

The mountain ranges and coastlines of Washington State have steep slopes, and they are susceptible to landslides triggered by intense rainstorms, rapid snow melts, earthquakes, and rivers and waves removing slope stability. Over a 30-year timespan (1984-2014 and includes State Route (SR) 530), a total of 28 deep-seated landslides caused 300 million dollars of damage and 45 deaths (DGER, 2015). During that same timeframe, ten storm events triggered shallow landslides and debris flows across the state, resulting in nine deaths (DGER, 2015). The loss of 43 people, due to the SR 530 complex reactivating and moving at a rate and distance unexpected to residents, highlighted the need for an inventory of the stateís landslides. With only 13% of the state mapped (Lombardo et al., 2015), the intention of this statewide inventory is to communicate hazards to citizens and decision makers. In order to compile an accurate and consistent landslide inventory, Washington needs to adopt a graphic information system (GIS) based mapping protocol. A mapping protocol provides consistency for measuring and recording information about landslides, including such information as the type of landslide, the material involved, and the size of the movement. The state of Oregon shares similar landslide problems as Washington, and it created a GIS-based mapping protocol designed to inform its residents, while also saving money and reducing costly hours in the field (Burns and Madin, 2009). In order to determine if the Oregon Department of Geology and Mineral Industries (DOGAMI) protocol, developed by Burns and Madin (2009), could serve as the basis for establishing Washingtonís protocol, I used the office-based DOGAMI protocol to map landslides along a 40-50 km (25-30 mile) shoreline in Thurston County, Washington. I then compared my results to the field-based landslide inventory created in 2009 by the Washington Division of Geology and Earth Resources (DGER) along this same shoreline. If the landslide area I mapped reasonably equaled the area of the DGER (2009) inventory, I would consider the DOGAMI protocol useful for Washington, too. Utilizing 1m resolution lidar flown for Thurston County in 2011 and a GIS platform, I mapped 36 landslide deposits and scarp flanks, covering a total area of 879,530 m2 (9,467,160 ft2). I also found 48 recent events within these deposits. With an exception of two slides, all of the movements occurred within the last fifty years. Along this same coastline, the DGER (2009) recorded 159 individual landslides and complexes, for a total area of 3,256,570 m2 (35,053,400 ft2). At a first glance it appears the DGER (2009) effort found a larger total number and total area of landslides. However, in addition to their field inventory, they digitized landslides previously mapped by other researchers, and they did not field confirm these landslides, which cover a total area of 2,093,860 m2 (22,538,150 ft2) (DGER, 2009). With this questionable landslide area removed and the toes and underwater landslides accounted for because I did not have a bathymetry dataset, my results are within 6,580 m2 (70,840 ft2) of the DGERís results. This similarity shows that the DOGAMI protocol provides a consistent and accurate approach to creating a landslide inventory. With a few additional modifications, I recommend that Washington State adopts the DOGAMI protocol. Acquiring additional 1m lidar and adopting a modified DOGAMI protocol poises the DGER to map the remaining 87% of the state, with an ultimate goal of informing citizens and decision makers of the locations and frequencies of landslide hazards on a user-friendly GIS platform.

The use of motivational interviews by nurses to promote health behaviors in adolescents: A scoping review protocol

Abstract The developmental changes during adolescence may affect subsequent risk for diseases and health-related behaviors. Traditionally, professionals assume that knowledge is sufficient for behavioral changes; however adolescents continue to engage in unhealthy behaviors despite clearly knowing what they should do and how to change. "What is lacking is the motivation to apply that knowledge". Motivational Interview (MI) may be taken as an essential tool in the provision of nursing care to adolescents, being itself a workspace with possible therapeutic effects. Objectives The objective of this scoping is to examine and map the use of MI by nurses in their clinical practice with adolescents to promote health behaviors. The review will focus on knowing what is the current extent of the use of nurse-led MI; which adolescent populations were included; in which contexts nurses use MI; which MI techniques/strategies have been used and what outcomes on health behaviors promotion have been reported. Methodology This scoping review will be informed by JBI methodology. The population of this study is adolescents aged 10 to 19 years participating in nurse-led MI. The concept of MI include MI done by personal or telephone call, with any number of sessions, brief interventions and other motivational interventions grounded but not limited to the principles described by Miller & Rollnick (2008). All geographical and all clinical practice contexts where nurses' undertake MI with adolescents such as hospitals, primary health care, health care centers, community or schools will be contemplated. English, Spanish and Portuguese published studies will be considered for inclusion. Results An initial limited search of MEDLINE and CINAHL was undertaken followed by analysis of the text words contained in the title and abstract, and of the index terms used to describe the concepts, synonyms (with truncations), MeSH Terms and Cinhal headings of this study. It was identified 5 synonyms for "Adolescents", 7 for "MI" and 2 for "nurse". A first search using the all 14 identified keywords and index terms was made at Medline (Title/Abstract) and brought up 125 articles. Other 16 databases referenced at the protocol will be searched to identify additional studies. Articles identified from the final search will be assessed for relevance to the review, based on information provided in the title and abstract. The full article will be retrieved for all studies that meet the inclusion criteria of the review. It is expected that findings from this Scoping Review provide needed information to nurses related to the use of MI to promote health behaviors in adolescents. Conclusions There is little knowledge of what works for whom (which adolescent subpopulation) under what circumstances (in which setting, for what problem) in relation to nurse-led MI. There is a need for scoping or mapping the nurse-led MI with adolescents to identify evidence gaps and to inform opportunities for future development in nursing practice. Moreover, information regarding implemented and evaluated interventions, techniques used, contexts of application and adolescents groups is dispersed in the literature which impedes the formulation of questions about the outcomes and effectiveness of those interventions. The practical implication of this mapping will be clarifying all these aspects.

The use of motivational interviews by nurses to promote health behaviors in adolescents: a scoping review protocol

Adolescents - defined as young people between 10 and 19 years of age1 - are, in general, a relatively healthy segment of the population.2 However, the developmental changes that take place during adolescence may affect their subsequent risk for diseases and for a variety of health-related behaviors. In fact, early onset of preventable health problems (e.g. obesity, malnutrition, STDs) and the engagement in health risk behaviors (e.g., sedentary life style, excessive alcohol consumption, unprotected sex) during adolescence, are likely to put them at greater risk for physical and mental health problems at a later stage in life. Moreover, health related problems and health risk behaviors may disrupt adolescents' physical and cognitive development and therefore may affect their ability to think and act in relation to decisions about their health in the future.1 In summary, health-related behaviors in adolescence, apart from their influence on the continuum of "health-disease", they also have the potential to influence future behaviors. In fact, several studies have shown that past behaviors are good predictors of future behaviors .3,4 Thus, promoting healthy practices during adolescence and taking measures to better protect young people from health risks are essential for the prevention of health problems in adulthood.5 According to the World Health Organization, the main problems affecting young people include mental health problems (such as behavioral disorders, eating disorders, suicide, anxiety or depression), the use of substances (illegal substances, alcohol and tobacco), interpersonal violence, nutrition (a proper nutrition consists of healthy eating habits and physical exercise), unintentional injuries (which are a leading cause of death and disability among young people, with road traffic injuries accounting for about 700 deaths per day), sexual and reproductive health (for example, risky sexual behaviors, early pregnancy and childbirth) and HIV (resulting from sexual transmission and drug injection).5,6 On the other hand, the number of children and youth with chronic health conditions has increased dramatically in the past four decades7 as larger numbers of chronically ill children survive beyond the age of 10.8 Despite the lack of data on adolescents' health making it difficult to determine the prevalence of chronic illnesses in this age group9, it is known that one in ten adolescents suffers from a chronic condition worldwide.10 In fact, national population based studies from Western countries show that 20-30% of teenagers have a chronic illness, defined as one that lasts longer than six months.8 The most prevalent chronic illness among adolescents is asthma and the one with the highest incidence is diabetes mellitus, particularly type II.9 Traditionally, healthcare professionals have been mainly investing in health education activities, through the transmission of knowledge with a view to creating habits, customs and behaviors, and promoting healthy lifestyles. However, empowering people does not only consist of giving them the right information11 , i.e. good information is not enough to cause people to make changes.12 The motivation or desire to change unhealthy behaviors and habits depends on many factors, namely intrinsic motivation, control over personal decisions, self-confidence and perception of effectiveness, personal ambivalence, and individualized assistance.12 Many professionals assume that supplying knowledge is sufficient for behavioral changes; however, even very good advice often fails to generate behavioral change. After all, people continue to engage in unhealthy behaviors despite clearly knowing what they should do and how to change. "What is lacking is the motivation to apply that knowledge".13, p.1233 In fact, behavioral change is a complex phenomenon with multiple determinants that also includes motivational variables. It is associated with ambivalent processes expressed in the dilemma between keeping the current status and moving on to new ways of acting. For example, telling adolescents that if they keep on engaging in a certain behavior, they are increasing the risk of developing a long-term condition such as cardiovascular disease, stroke or diabetes is rarely enough to trigger the desired behavioral change; people are more likely to change when they believe that the change is really effective and that they are able to implement it.12 Therefore, it is essential to provide specific training for "healthcare professionals to master motivational techniques, avoid confrontation with the users, and facilitate behavioral changes".14 In this context, motivating patients to make behavioral changes is also an important nursing task where change in lifestyle is a major element of patients' treatment and preventive interventions.15 One of the nurse's goals is to help improve a patient's health or help them to manage existing health conditions. Once nurses are in a position where they have to focus on accomplishing tasks and telling patients what needs to be accomplished16, the role of the nurse is expanding even more into the use of motivational strategies.17 MI is bringing nurses back to therapeutic communication and moving them closer to successful health promotion and disease management, by promoting behavior change and empowering their patients. As the nursing profession evolves, MI is seen as a challenge and the basis of nurse's interactions with individuals, families and communities.16, 17 In the same way, MI may be taken as an essential tool in the provision of nursing care to adolescents, being itself a workspace with possible therapeutic effects regarding problems, clarification of doubts, and development of skills.18 In fact, MI may be particularly applicable in work with adolescents because of their specific developmental stage. Adolescents attempt to establish their own autonomy and identity while struggling with social interactions and moral issues, which leads to ambivalence.19 Consistent with the developmental challenges during adolescence, "MI explicitly honors autonomy, people's right and irrevocable ability to decide about their own behavior"20 while allowing the person to explore possibilities for change of risky or maladaptive behaviours.19 MI can be defined as a directive, client-centred counselling style for eliciting behavior change by helping clients to explore and resolve ambivalence. It is most centrally defined not by technique but by its spirit as a facilitative style of interpersonal relationship.21 It is a set of strategies and techniques widely used in clinical practice based on the transtheoretical model of change. The Stages of Change model describes five stages of readiness—precontemplation, contemplation, preparation, action, and maintenance—and provides a framework for understanding behavior change.22 The MI has been widely tested and applied in different areas, such as modification of addictive behaviors, interventions with offenders in the context of justice, eating disorders, promotion of therapeutic adherence among chronic patients, promotion of learning in school settings or intervention with adolescents at risk.18,23 In general, clinical practice has been adopting the perspective of motivation as something relatively immutable, i.e., the adolescent is either motivated for change/treatment and, in these conditions, the professional's role is to help him/her, or the adolescent is not motivated and then change/treatment is not feasible. Alternatively the theoretical model underlying the MI technique postulates that the individual's adherence to change/treatment depends on his/her motivation, which can change throughout the therapeutic intervention. As several studies found positive results for effects of MI24-26 and its use by health professionals is encouraged23,27 nurses may play an important role in patients' process of change. As nurses have a crucial role in clinical contexts, they can facilitate the process of ending risk behaviors and/or adopting positive health behaviors through some motivational techniques, namely with adolescents. A considerable number of systematic reviews about MI already exist pointing to some benefits of its use in the treatment of a broad range of behavioral problems and diseases.13,28,29 Some of the current reviews focus on examining the effectiveness of MI for adolescents with diverse health risks/problems 30-32. However, to date there are no reviews that present and assess the evidence for the use of nurse-led MI in adolescents. Therefore, we have little knowledge of what works for whom (which adolescent subpopulation) under what circumstances (in which setting, for what problem) in relation to motivational interviewing by nurses. There is a clear need for scoping or mapping the use of MI by nurses with adolescents to identify evidence gaps and to inform opportunities for future development in nursing practice. On the other hand, information regarding nurse-led implemented and evaluated interventions, techniques and/or strategies used, contexts of application and adolescents subpopulation groups is dispersed in the literature33-36 which impedes the formulation of precise questions about the effectiveness of those interventions conducted by nurses and therefore the realization of a systematic review. In other words, it is known that different kind of motivational interventions have been implemented in different contexts by nurses, however does not exist a map about all the motivational techniques and/or strategies used. Furthermore the literature does not clarify which is the role of nurses at cross professional motivational intervention implemented programs and finally the outcomes and evaluation of interventions are unclear. Thus, the practical implication of this mapping will be clarifying all these aspects. Without this clarification is not possible to proceed to the realization of a systematic review about the effectiveness of the use of motivational interviews by nurses to promote health behaviors in adolescents, in a particular context and/or health risk behavior; or regarding the effectiveness of certain technique and/or strategy of MI. Consequently, there are important questions about the nature of the evidence in this area that need to be answered before formulating a precise question of effectiveness. This scoping review aims to respond to these questions. An initial search of the JBI Database of Systematic Reviews & Implementation Reports, Cochrane Database of Systematic Reviews, , Database of promoting health effectiveness reviews (DoPHER), The Campbell Library, Medline and CINAHL, has revealed that currently there is no Scoping Review (published or in progress) on the subject. In this context, this scoping review will examine and map the published and unpublished research around the use of MI by nurses implemented and evaluated to promote health behaviors in adolescents; to establish its current extent, range and nature and identify its feasibility, outcomes and gaps in the evidence defining research priorities in this field. This scoping review will be informed by the JBI methodology37 that suggests a five stage methodological framework for conducting scoping reviews which includes: identifying the research question, searching for relevant studies, selecting studies, charting data, collating, summarizing and reporting the results.

The use of motivational interviews by nurses to promote health behaviors in adolescents: A scoping review protocol

Abstract The developmental changes during adolescence may affect subsequent risk for diseases and health-related behaviors. Motivational Interview (MI) may be taken as an essential tool in the provision of nursing care to adolescents, being itself a workspace with possible therapeutic effects. At this context, it is important to examine and map the use of MI by nurses in their clinical practice with adolescents to promote health behaviors. Objetives A scoping review has been currently undertaking to find out what is the current extent of the use of nurse-led MI; which adolescent populations were included; in which contexts nurses use MI; which MI techniques/strategies have been used and what outcomes on health behaviors promotion have been reported. The first task was to develop a search strategy to identify relevant studies for this review. It is described here the experience in constructing a search strategy for this review. Methodology This scoping review will be informed by JBI methodology. An initial search of MEDLINE and CINAHL was undertaken. An analysis of the text words contained in the title and abstract, and of the index terms used to describe the searched articles were retrieved and these were then used to construct a search strategy for use in Medline and Cinahl. Results Analysis of the text words and the index terms generated 19 keywords: It was identified 5 synonyms for "Adolescents", 12 for "MI" and 2 for "nurse". A new research formula was designed using the text words identified. Adolescen*[Title/Abstract]) OR Younger*[Title/Abstract]) OR Youth*[Title/Abstract]) OR Teen*[Title/Abstract]) OR Adolescent[MeSH Terms])) AND (((((((((((("Motivational interview"[Title/Abstract]) OR "Motivational intervention"[Title/Abstract]) OR "Motivational interviews"[Title/Abstract]) OR "Motivational interventions"[Title/Abstract]) OR "motivational interviewing"[Title/Abstract]) OR "motivational counseling"[Title/Abstract]) OR "motivational support"[Title/Abstract]) OR "Motivational enhancement"[Title/Abstract]) OR "Brief intervention"[Title/Abstract]) OR "Brief interventions"[Title/Abstract]) OR Motivational Interviewing[MeSH Terms]) OR Directive Counseling[MeSH Terms])) AND ((nurs*[Text Word]) OR Nurses[MeSH Terms]). Limiters - Language: English, Portuguese, Spanish. In MEDLINE this research formula generated 125 results. Other 16 databases referenced at the protocol will be searched to identify additional studies. Articles identified from the final search will be assessed for relevance to the review, based on information provided in the title and abstract. The full article will be retrieved for all studies that meet the inclusion criteria of the review. Conclusions It was presented here the initial results of this search. Next steps of this study will be to develop and refine the search strategy for use in other databases. It is expected that findings from this Scoping Review provide needed information to nurses related to the use of MI to promote health behaviors in adolescents and inform opportunities for future development in nursing practice.

An Efficient Protocol For The Generation Of Monocyte Derived Dendritic Cells Using Serum-free Media For Clinical Applications In Post Remission Aml Patients.

Protocols for the generation of dendritic cells (DCs) using serum as a supplementation of culture media leads to reactions due to animal proteins and disease transmissions. Several types of serum-free media (SFM), based on good manufacture practices (GMP), have recently been used and seem to be a viable option. The aim of this study was to evaluate the results of the differentiation, maturation, and function of DCs from Acute Myeloid Leukemia patients (AML), generated in SFM and medium supplemented with autologous serum (AS). DCs were analyzed by phenotype characteristics, viability, and functionality. The results showed the possibility of generating viable DCs in all the conditions tested. In patients, the X-VIVO 15 medium was more efficient than the other media tested in the generation of DCs producing IL-12p70 (p=0.05). Moreover, the presence of AS led to a significant increase of IL-10 by DCs as compared with CellGro (p=0.05) and X-Vivo15 (p=0.05) media, both in patients and donors. We concluded that SFM was efficient in the production of DCs for immunotherapy in AML patients. However, the use of AS appears to interfere with the functional capacity of the generated DCs.

A Metabolomic Protocol For Plant Systematics By Matrix-assisted Laser-desorption/ionization Time-of Flight Mass Spectrometry.

Matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS) has been widely used for the identification and classification of microorganisms based on their proteomic fingerprints. However, the use of MALDI-TOF MS in plant research has been very limited. In the present study, a first protocol is proposed for metabolic fingerprinting by MALDI-TOF MS using three different MALDI matrices with subsequent multivariate data analysis by in-house algorithms implemented in the R environment for the taxonomic classification of plants from different genera, families and orders. By merging the data acquired with different matrices, different ionization modes and using careful algorithms and parameter selection, we demonstrate that a close taxonomic classification can be achieved based on plant metabolic fingerprints, with 92% similarity to the taxonomic classifications found in literature. The present work therefore highlights the great potential of applying MALDI-TOF MS for the taxonomic classification of plants and, furthermore, provides a preliminary foundation for future research.

Protocol for DNA extraction of Cryptosporidium spp. oocysts in fecal samples

Molecular characterization of Cryptosporidium spp.oocysts in clinical samples is useful for public health since it allows the study of sources of contamination as well as the transmission in different geographical regions. Although widely used in developed countries, in Brazil it is restricted to academic studies, mostly using commercial kits for the extraction of genomic DNA, or in collaboration with external reference centers, rendering the method expensive and limited. The study proposes the application of the modifications recently introduced in the method improving feasibility with lower cost. This method was efficient for clinical samples preserved at -20 °C for up to six years and the low number of oocysts may be overcomed by repetitions of extraction.

Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium

A morphological and cell culture study from nasal mucosa of dogs was performed in order to establish a protocol to obtain a cell population committed to neuronal lineage, as a proposal for the treatment of traumatic and degenerative lesions in these animals, so that in the future these results could be applied to the human species. Twelve mongrel dogs of 60-day aged pregnancy were collected from urban pound dogs in São Paulo. Tissue from cribriform ethmoidal lamina of the fetuses was collected at necropsy under sterile conditions around 1h to 2h postmortem by uterine sections and sections from the fetal regions described above. Isolated cells of this tissue were added in DMEM/F-12 medium under standard conditions of incubation (5% CO², >37ºC). Cell culture based on isolated cells from biopsies of the olfactory epithelium showed rapid growth when cultured for 24 hours, showing phase-bright sphere cells found floating around the fragments, attached on culture flasks. After 20 days, a specific type of cells, predominantly ellipsoids or fusiform cells was characterized in vitro. The indirect immunofluorescence examination showed cells expressing markers of neuronal precursors (GFAP, neurofilament, oligodendrocyte, and III â-tubulin). The cell proliferation index showed Ki67 immunostaining with a trend to label cell groups throughout the apical region, while PCNA immunostaining label predominantly cell groups lying above the basal lamina. The transmission electron microscopy from the olfactory epithelium of dogs revealed cells with electron-dense cytoplasm and preserving the same distribution as those of positive cell staining for PCNA. Metabolic activity was confirmed by presence of euchromatin in the greatest part of cells. All these aspects give subsidies to support the hypothesis about resident progenitor cells among the basal cells of the olfactory epithelium, committed to renewal of these cell populations, especially neurons.

A rational protocol for the successful crystallization of l-amino-acid oxidase from Bothrops atrox

Despite the valuable contributions of robotics and high-throughput approaches to protein crystallization, the role of an experienced crystallographer in the evaluation and rationalization of a crystallization process is still crucial to obtaining crystals suitable for X-ray diffraction measurements. In this work, the difficult task of crystallizing the flavoenzyme l-amino-acid oxidase purified from Bothrops atrox snake venom was overcome by the development of a protocol that first required the identification of a non-amorphous precipitate as a promising crystallization condition followed by the implementation of a methodology that combined crystallization in the presence of oil and seeding techniques. Crystals were obtained and a complete data set was collected to 2.3 A resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 73.64, b = 123.92, c = 105.08 A, beta = 96.03 degrees. There were four protein subunits in the asymmetric unit, which gave a Matthews coefficient V (M) of 2.12 A3 Da-1, corresponding to 42% solvent content. The structure has been solved by molecular-replacement techniques.

Severity of Oral Mucositis in Patients Undergoing Hematopoietic Cell Transplantation and an Oral Laser Phototherapy Protocol: A Survey of 30 Patients

Background Data and Objective: Oral mucositis (OM) is one of the worst cytotoxic effects of chemotherapy and radiotherapy in patients undergoing hematopoietic cell transplantation (HCT), and it causes severe morbidity. Laser phototherapy has been considered as an alternative therapy for prevention and treatment of OM. The aim of this study was to describe the incidence and severity of OM in HCT patients subjected to laser phototherapy, and to discuss its effect on the oral mucosa. Patients and Methods: Information concerning patient age and gender, type of basic disease, conditioning regimen, type of transplant, absence or presence of pain related to the oral cavity, OM grade, and adverse reactions or unusual events were collected from 30 patients undergoing HCT (allogeneic or autologous). These patients were given oral laser phototherapy with a InGaAIP laser (660 nm and 40 mW) daily. The data were tabulated and their frequency expressed as percentages. Results: In the analysis of those with OM, it was observed that 33.4% exhibited grade I, 40% grade II, 23.3% grade III, and 3.3% grade IV disease. On the most critical post-HCT days (D+5 and D+8), it was observed that 63.3% of patients had grade I and 33.3% had grade II disease; no patients had grade III or IV disease in this period. This severity of OM was similar to that seen in other studies of laser phototherapy and OM. Conclusion: The low grades of OM observed in this survey show the beneficial effects of laser phototherapy, but randomized clinical trials are necessary to confirm these findings.