La imagen radioisotópica: una herramienta eficaz en la terapia regenerativa / Radioisotopic image: an efficient tool in regenerative therapy

La terapia regenerativa constituye hoy una prometedora alternativa de tratamiento para muchas enfermedades. En este trabajo se describe la utilidad de las tecnologías de imagen radioisotópica en la evaluación de la seguridad y eficacia de la terapia con células madre. Para ello se exponen algunos de los fenómenos que condujeron al desarrollo de la tecnología de imagen en medicina nuclear, así como las características generales de la tomografía computarizada por emisión de fotón único y por emisión de positrones. También se reseña la aplicación de las imágenes radioisotópicas en la evaluación de los cambios de biodistribución, flujo y perfusión en las zonas anatómicas afectadas por distintas enfermedades en las especialidades de angiología, ortopedia y cardiología, como resultado de los efectos regenerativos y proliferativos inducidos por las células madre. Se citan los radiofármacos y moléculas marcadas que se usan en la evaluación de la angiogénesis y la linfangiogénesis resultantes de la terapia celular y los marcadores radioisotópicos que permiten el rastreo de las células madre trasplantadas(AU)

Regenerative therapy is nowadays a promising alternative treatment for many diseases. The utility of radioisotope image technologies in assessing of the efficiency and security of stem cell therapies is described. We present some of the facts that led to development of the nuclear medicine image technology, as well as the general characteristics of single photon emission computed tomography and positron emission tomography. It was also included application of radioisotope images in the evaluation of biodistribution changes, flow and perfusion in anatomical affected zones by different diseases, in the fields of angiology, orthopedia and cardiology, as a result of the regenerative and proliferative effects induced by stem cells transplant. We mention the radiopharmaceuticals and labelled molecules used in the evaluation of angiogenesis and lymph angiogenesis resulting of cellular therapy and the radioisotopes markers that allow tracking of transplanted stem cells(AU)

El conteo automático de reticulocitos: una herramienta de uso diagnóstico, clínico e investigativo / Automated reticulocyte count: a tool for diagnostic, clinical and research use

Se revisan los antecedentes históricos del conteo de reticulocitos como determinación indispensable en el laboratorio de hematología para la evaluación de la actividad eritropoyética durante la clasificación, diagnóstico y monitoreo de la respuesta terapéutica en distintos trastornos y situaciones clínicas, principalmente en casos de anemias. Se describe el tránsito del tradicional método de conteo de reticulocitos manual al método automatizado y la integración de los parámetros reticulocitarios al hemograma automatizado actual; además, se analizan las desventajas del método de recuento manual y las ventajas del método automatizado, así como los principios de detección en que se basa el conteo electrónico de reticulocitos. Con relación a los parámetros reticulocitarios, se describe su medición, cálculo y unidades de medida; también se resalta la importancia de la fracción de reticulocitos inmaduros y del contenido de hemoglobina reticulocitaria como variables de mayor uso clínico e investigativo en la evaluación de la respuesta medular ante diversos trastornos clínicos y protocolos terapéuticos. Por último, se alude a la necesidad del conocimiento y empleo de las variables reticulocitarias en la práctica clínica de rutina por parte de los clínicos y especialistas en hematología.

The historical background of reticulocyte count is reviewed as an essential determination in the laboratory of hematology for the evaluation of erythropoietic activity during classification, diagnosis and monitoring of therapeutic response of different conditions and clinical situations are also reviewed, especially in anemia The transition from traditional manual reticulocyte counting method to automated method and integration of the reticulocyte parameters to current automated complete blood count are described. The disadvantages of manual method and the advantages of automated methods are cited, as well as detection principles in which electronic reticulocyte count is based. Regarding reticulocyte parameters, measurement, calculation and units are described. The importance of immature reticulocyte fraction and reticulocyte hemoglobin content as variables most clinical and research use in evaluating bone marrow response to various clinical disorders and therapeutic protocols are highlighted. Finally, the need for knowledge and use of reticulocyte variables in routine clinical practice by clinicians and hematologist is referred.

Ética y protección radiológica en la medicina nuclear en hematología / Ethics and radiological protection in nuclear medicine in hematology

El campo de utilización cada vez más amplio de las sustancias radiactivas en la medicina exige el manejo y la aplicación de los principios de la ética en una relación muy estrecha con la protección radiológica (PR). Existe una necesidad insoslayable de llevar la perspectiva de la ética en el uso de las radiaciones ionizantes y de la PR del paciente a la práctica de la Medicina Nuclear en Hematología (MNH). Esto no debe asumirse como un proceso de simple adopción, sino como la aplicación creativa de sus principios. Será de gran utilidad continuar incluyendo en la docencia que se imparte a los profesionales de las diversas especialidades que emplean la medicina nuclear como herramienta diagnóstica o terapéutica, una formación científica basada en los principios de la ética sumados a elementos de la PR. El colectivo multidisciplinario dedicado a la MNH tiene una responsabilidad de índole moral y ética en la garantía del uso adecuado de las sustancias radiactivas en la técnica médica y el desarrollo científico de la especialidad. La siguiente revisión del tema pretende contribuir al fomento de principios éticos en los especialistas de hematología durante su quehacer diario. Con la aplicación de la ética en el contexto de la MNH, perfeccionaremos la gestión de PR en el manejo de los pacientes, contribuiremos a la mejor atención a estos y elevaremos la calidad de nuestro trabajo...

The increasingly width of the field of radioactive substances use in medicine demands the managing and the application of ethics principles in a very close relation to radiological protection. There is an unavoidable need to take ethics perspective in the use of ionizing radiations and the radiological protection of the patient to the practice of Nuclear Medicine in Hematology (NMH). This must not be assumed as a process of simple adoption, but as the creative application of his principles. It will be highly useful to continue including in the teaching programs aimed to professionals who use nuclear medicine as a diagnostic or therapeutic tool, a scientific formation based on the ethics principles including the elements of radiological protection. The multidisciplinary group dedicated to NMH has a moral and ethical responsibility to guarantee the appropriate use of radioactive substances in the medical technology and scientific development of this specialty. The following review on the subject wishes to contribute to the promotion of ethical principles in hematology specialists during their daily work. With the application of ethics in the context of NMH, we will also improve the radiological protection of our patients which will contribute to their best care and will increase the quality of our work...

Ética y protección radiológica en la medicina nuclear en hematología / Ethics and radiological protection in nuclear medicine in hematology

El campo de utilización cada vez más amplio de las sustancias radiactivas en la medicina exige el manejo y la aplicación de los principios de la ética en una relación muy estrecha con la protección radiológica (PR). Existe una necesidad insoslayable de llevar la perspectiva de la ética en el uso de las radiaciones ionizantes y de la PR del paciente a la práctica de la Medicina Nuclear en Hematología (MNH). Esto no debe asumirse como un proceso de simple adopción, sino como la aplicación creativa de sus principios. Será de gran utilidad continuar incluyendo en la docencia que se imparte a los profesionales de las diversas especialidades que emplean la medicina nuclear como herramienta diagnóstica o terapéutica, una formación científica basada en los principios de la ética sumados a elementos de la PR. El colectivo multidisciplinario dedicado a la MNH tiene una responsabilidad de índole moral y ética en la garantía del uso adecuado de las sustancias radiactivas en la técnica médica y el desarrollo científico de la especialidad. La siguiente revisión del tema pretende contribuir al fomento de principios éticos en los especialistas de hematología durante su quehacer diario. Con la aplicación de la ética en el contexto de la MNH, perfeccionaremos la gestión de PR en el manejo de los pacientes, contribuiremos a la mejor atención a estos y elevaremos la calidad de nuestro trabajo(AU)

The increasingly width of the field of radioactive substances use in medicine demands the managing and the application of ethics principles in a very close relation to radiological protection. There is an unavoidable need to take ethics perspective in the use of ionizing radiations and the radiological protection of the patient to the practice of Nuclear Medicine in Hematology (NMH). This must not be assumed as a process of simple adoption, but as the creative application of his principles. It will be highly useful to continue including in the teaching programs aimed to professionals who use nuclear medicine as a diagnostic or therapeutic tool, a scientific formation based on the ethics principles including the elements of radiological protection. The multidisciplinary group dedicated to NMH has a moral and ethical responsibility to guarantee the appropriate use of radioactive substances in the medical technology and scientific development of this specialty. The following review on the subject wishes to contribute to the promotion of ethical principles in hematology specialists during their daily work. With the application of ethics in the context of NMH, we will also improve the radiological protection of our patients which will contribute to their best care and will increase the quality of our work(AU)

El hemograma: nueva clasificación y perspectivas / Hemogram: new classification and perspectives

A través del tiempo, el surgimiento y desarrollo de nuevos métodos analíticos se han visto unidos a la introducción de nuevas tecnologías y a su incorporación al trabajo diagnóstico e investigativo del laboratorio clínico. En los últimos años, los indicadores de exactitud y precisión de los procedimientos en el laboratorio de hematología se han elevado extraordinariamente gracias a la llegada de los sistemas automatizados de conteo y caracterización de células sanguíneas.Desde principios de la segunda mitad del pasado siglo, la continúa fabricación de diversos modelos de contadores hematológicos por parte de las compañías proveedoras y firmas comerciales, ha nutrido el mercado internacional de distintas series de estos equipos. La combinación de principios de detección tales como la impedancia eléctrica, la radiofrecuencia, las medidas de dispersión y absorción de la luz halógena o láser en diversos ángulos y la citometría de flujo, como bases del conteo y caracterización de las poblaciones celulares hemáticas, ha hecho posible el surgimiento de autoanalizadores hematológicos de mayor costo y complejidad.Como fruto de los avances de esta tecnología y su aplicación al laboratorio de hematología, el hemograma o biometría hemática, como indicación de primera línea en la evaluación clínica de los desórdenes y respuestas del sistema hematopoyético, es hoy día una de las pruebas más accesibles y solicitadas al laboratorio clínico.1,2Esta técnica ha sido objeto de infinidad de variaciones en aspectos tales como la forma automatizada o manual de su realización, el número de parámetros que la componen, la manera de interpretarlos y la elevación de los indicadores de exactitud y precisión de sus..

El hemograma: nueva clasificación y perspectivas / Hemogram: new classification and perspectives

A través del tiempo, el surgimiento y desarrollo de nuevos métodos analíticos se han visto unidos a la introducción de nuevas tecnologías y a su incorporación al trabajo diagnóstico e investigativo del laboratorio clínico. En los últimos años, los indicadores de exactitud y precisión de los procedimientos en el laboratorio de hematología se han elevado extraordinariamente gracias a la llegada de los sistemas automatizados de conteo y caracterización de células sanguíneas.Desde principios de la segunda mitad del pasado siglo, la continúa fabricación de diversos modelos de contadores hematológicos por parte de las compañías proveedoras y firmas comerciales, ha nutrido el mercado internacional de distintas series de estos equipos. La combinación de principios de detección tales como la impedancia eléctrica, la radiofrecuencia, las medidas de dispersión y absorción de la luz halógena o láser en diversos ángulos y la citometría de flujo, como bases del conteo y caracterización de las poblaciones celulares hemáticas, ha hecho posible el surgimiento de autoanalizadores hematológicos de mayor costo y complejidad.Como fruto de los avances de esta tecnología y su aplicación al laboratorio de hematología, el hemograma o biometría hemática, como indicación de primera línea en la evaluación clínica de los desórdenes y respuestas del sistema hematopoyético, es hoy día una de las pruebas más accesibles y solicitadas al laboratorio clínico.1,2Esta técnica ha sido objeto de infinidad de variaciones en aspectos tales como la forma automatizada o manual de su realización, el número de parámetros que la componen, la manera de interpretarlos y la elevación de los indicadores de exactitud y precisión de sus..(AU)

Prediction of digestibility and energy concentration of winter pasture forage and herbage of low-input grassland--a comparison of methods.

Regarding the estimation of the energy concentration or digestibility of herb-dominated forage and plant samples from winter pastures, it could be expected that the estimation is only reliable when in vitro methods with rumen fluid as inoculum (= gas production techniques) are used. For the verification of this thesis based on logical reflections, an in vitro-method with rumen fluid added as inoculum, as well as chemical, and enzymatic methods were applied under consideration of existing estimating functions. As a possible reason for the observed divergence of the methods, effects of fungal infections or, respectively, secondary compounds in herbs are discussed. At the present state of knowledge, it is adequate to estimate the energy concentration in vitro by gas tests, as far as fattening types like suckler cows and beef cattle are concerned, maybe in contrast to the forage evaluation for dairy cows.

Effects of the botanical composition and weather conditions on mycotoxins in winter forage from grassland.

The paper focuses on possible effects of the botanical composition and weather conditions on the extend of zearalenone and ochratoxin A concentrations in various grass swards in winter pasture systems. Zearalenone is only detectable in pure stands ofLolium perenne or inLolio-Cynosuretum plant communities, respectively. The occurrence of ochratoxin A is more frequent and less specific concerning the botanical composition. This mycotoxin was found in both,Lolium perenne andFestuca arundinacea in varying years. The incidence of ochratoxin A depends on year and is apparently related to the weather conditions in autumn and winter. There was no evidence that particular locatio8/22/2008 12:57PMns have a higher or a lower risk for high ochratoxin A or zearalenone concentrations than others. Peak values in individual swards are not intermittent over the years.

Light scattering in normal and cataractous lenses of farmed Atlantic salmon (Salmo salar): a slit lamp and Scheimpflug photographic study.

To investigate normal light scattering and cataract formation, the anterior eye segments of farmed Atlantic salmon (Salmo salar) reared in fresh water and sea water were documented in vivo for the first time with a Topcon SL-45 Scheimpflug camera. A total of 40 fish from the fresh-water-rearing period, obtained from 2 groups of identical age but showing a different growth rate, and 24 fish from the sea-water-rearing period, sampled from 2 groups with identical age but being fed different food brands, were included in this study. The fish were anaesthetized before examination. Due to the naturally wide pupil, no mydriatic compound was applied. All fish were removed from the water for photography, which was performed for each eye in 0 degrees = vertical slit position. Images were recorded on Kodak Tmax 400 black-and-white film. Microdensitometric image analysis of all negatives was performed using a Joyce-Loebl online microdensitometer. In spite of the virtual absence of an anterior chamber gap between cornea and lens and very little light scattering in the normal fish lens, a small number of distinct layers could be reproducibly identified in the lens. While there was little abnormal light scattering which could point to cataract development in young fish from the fresh water period, the evaluation of the lenses from the 2 sea water groups showed the presence of specific forms of cataract especially in the cortical and supranuclear layers. There were significant differences between the groups fed different food brands at the sea water site. In conclusion, Scheimpflug photography proved to be applicable to eye research in fish in vivo. It is suggested that this method should be employed for reproducible documentation as an extension to slit lamp monitoring in experimental research to reveal causative factors for cataracts in farmed fish.

[Image analysis and Sheimpflug photography of anterior segment of the eye--a review]. / Optische Schnittbild-Vermessung des vorderen Augenabschnittes nach Scheimpflug: Möglichkeiten und Grenzen--eine Ubersicht.

BACKGROUND: Light scattering properties of the cornea and lens change in relation to age. This process is influenced by noxious factors which may induce the formation of cataracts. Four different camera types based on the Scheimpflug principle have been developed for documentation and evaluation of light scattering in the eye. Topcon SL-45. Zeiss SLC, Oxford CASE 2000, Nidek EAS 1000. Only the SL-45 and EAS 1000, however, have found a wider application in clinical and experimental studies. MATERIAL AND METHODS: Documentation of the eye with a Scheimpflug system is performed in a dark room after mydriasis induction. The rotating axis of the camera is aligned to the optical axis of the eye with fixation devices. Four meridians have been found appropriate for routine documentation. Image analysis is based on peak height and distance evaluation of 2 layers in the cornea and 10 layers in the lens. Standardization devices allow correction of technical differences in the recording process. RESULTS: Evaluation of the ageing properties of the lens has evidenced typical density developments for each individual layer, the cortex increasing in density earlier than the nucleus. Cataract classification using a Scheimpflug camera provides an objective characterization of cataract type and progression. The outcome of surgical procedures changing the refractive power of the eye can be evaluated with a Scheimpflug camera as well. Its application in studies dealing with the ocular effects of Quinolones and HMG-CoA-reductase inhibitors, has demonstrated that Scheimpflug photography is the method of choice for establishment of a safety profile for a new drug. CONCLUSIONS: Scheimpflug photography has proven to be ideal for objective documentation of the anterior eye segment in human and animal eyes, although both documentation and image analysis demand technical expertise to be reproducible.

A new screen for protein interactions reveals that the Saccharomyces cerevisiae high mobility group proteins Nhp6A/B are involved in the regulation of the GAL1 promoter.

The split-ubiquitin assay detects protein interactions in vivo. To identify proteins interacting with Gal4p and Tup1p, two transcriptional regulators, we converted the split-ubiquitin assay into a generally applicable screen for binding partners of specific proteins in vivo. A library of genomic Saccharomyces cerevisiae DNA fragments fused to the N-terminal half of ubiquitin was constructed and transformed into yeast strains carrying either Gal4p or Tup1p as a bait. Both proteins were C-terminally extended by the C-terminal half of ubiquitin followed by a modified Ura3p with an arginine in position 1, a destabilizing residue in the N-end rule pathway. The bait fusion protein alone is stable and enzymatically active. However, upon interaction with its prey, a native-like ubiquitin is reconstituted. RUra3p is then cleaved off by the ubiquitin-specific proteases and rapidly degraded by the N-end rule pathway. In both screens, Nhp6B was identified as a protein in close proximity to Gal4p as well as to Tup1p. Direct interaction between either protein and Nhp6B was confirmed by coprecipitation assays. Genetic analysis revealed that Nhp6B, a member of the HMG1 family of DNA-binding proteins, can influence transcriptional activation as well as repression at a specific locus in the chromosome of the yeast S. cerevisiae.

The AT-rich region between -54 to -66 is important for the promoter activity of interleukin-10 in Epstein-Barr virus positive Burkitt's lymphoma cells.

IL-10 is an important regulatory cytokine. The recent characterization of the 5'-flanking region of IL-10 led to the identification of the promoter region. The infection of B cells with EBV induces IL-10 production which may contribute to EBV-induced transformation. In the present report, IL-10 promoter elements involved in the constitutive expression of IL-10 in EBV-positive lymphoma cells are described. The AT-rich region between -54/-66 from the transcriptional start site was found to be important for IL-10-promoter activity in BL36. A point mutation at position -60 (T/A) was associated with over 90% reduction of luciferase activity in the cell lines BL36 and BL74. The conversion of A/T at position -57 led to enhanced promoter activity. In addition the AT-rich region could serve as an enhancer for the beta-globin basic promoter. In BJAB cells (EBV-negative), sequences between -205/-139 rather than the AT-rich region were involved in IL-10 promoter regulation. This underlines the importance of the AT-rich region for EBV-associated IL-10 promoter regulation. Our results further the understanding of how the IL-10 gene could be regulated in B cell lymphomas.

Imaging of the spatial density distribution on the capsule of the lens with Scheimpflug photography.

The density characteristics of the anterior and posterior surfaces of the lens, although of primary importance for its optical quality, are difficult to determine. Scheimpflug photography, however, allows distinct density measurements from the optical axis towards the periphery with the only limitation of a sufficient mydriasis, so that a 3-dimensional scatter profile of both capsular surfaces can be calculated via multilinear densitometric evaluation of a sufficient number of meridians. On this methodical basis, we analysed Scheimpflug images recorded with the Topcon SL-45 on Kodak T-Max 400 film from 25 patients in an age range between 19 and 63 years. Images from the 0 degree + 45 degree meridians were scanned with multilinear microdensitometry and the data computed to get a density profile. In general, most scattering profiles show a marked interindividual variability. Nevertheless, a general trend can be established between the periphery and the center of the anterior and posterior capsules: (a) the anterior capsule generally has a higher density than the posterior; (b) in most cases, the center of the posterior capsule has a higher density than the periphery; (c) the anterior capsule has a more uniform spatial density distribution in all cases. From the cases evaluated, it can be concluded that the density of the peripheral posterior capsule is more importantly influenced by shading effects of the iris, so that geometrical correction is needed.

[Objectified measurement of eye lens transparency in elderly probands. Results of a Scheimpflug photography study over the course of three and a half years]. / Objektivierte Messung der Augenlinsentransparenz in einer älteren Probandenpopulation. Ergebnisse einer Scheimpflug-Photographischen Studie über einen Zeitraum von 3,5 Jahren.

Lens density has been followed-up in 32 healthy volunteers (mean age 65 years) over a period of 3.5 years. The negatives of Scheimpflug-photos (Topcon SL-45) were evaluated by linear microdensitometry in three measure planes (Joyce Loebl 3 CS). Correlation coefficients, curves of regression as well as calculations of the percentage of density change made obvious, that a minimal increase in lens density occurred in the anterior cortical region.

Objectified measurements of eye lens transparency in a volunteer group of advanced age, carried out over a period of 3.5 years. Results of a Scheimpflug-photographic study.

The present study deals with a prospective investigation on the optical density of the eye lenses of 32 volunteers with initially normal eyes (mean age 65 years) over a period of 3.5 years. The negatives of the Scheimpflug photographs (Topcon SL-45) were evaluated by linear densitometry in 3 measure planes (Joyce Loebl 3CS). The evaluation of the coefficient of correlation, the representation of the regression straight line as well as the evaluation of the percent changes in lens density showed that during the observation period of 3.5 years a minimum increase in density took place mainly in the anterior cortex.

Comparison of the Nidek EAS 1000 system and the Topcon SL-45 in clinical application.

Some years ago, Nidek developed a new imaging system for the anterior eye segment, which offers the possibility of recording Scheimpflug and retroillumination images. The system consists of 2 different technical units, a camera unit for image recording and a computer unit for storage, system operation and image analysis. To evaluate the clinical use of the system and its reproducibility, a study with 31 volunteers/patients was performed that were photographed with the Nidek EAS 1000 at the meridians 0 degree, 45 degrees and 180 degrees. Two examinations with a 1-week interval were performed. In addition, a direct comparison of the EAS 1000 with the Topcon SL-45 was carried out with 32 volunteers that were photographed at 0 degree and 45 degrees; a retroillumination photo was additionally recorded. The video images were evaluated with the computer software, the SL-45 negatives were standardly measured with a Joyce-Loebl densitometer. The handling of the EAS 1000 in clinical use was found to be easy and comfortable for the patient, as the alignment is done with infrared light. Difficulties occur only with the retroillumination photography in that the infrared light for imaging may outshine the red fixation light during exposure. However, shadowing problems due to the eyelashes occur in oblique slit positions, especially in those patients with a pronounced front head. In studying the reproducibility, the mean values of the individual coefficients of variation for light scattering (density) were in a range between 3.6 and 5.06%, but more than 35% of the single values show a variation coefficient above 5% (maximum 20.3%).(ABSTRACT TRUNCATED AT 250 WORDS)

Reproducibility studies with the Zeiss SLC system and animal cataract models.

Scheimpflug photography has become the basic method for documentation of lens transparency changes in clinical and experimental ophthalmology, due to its high reproducibility. In animal cataract studies, the reproducibility is strongly influenced by the handling characteristics of the camera used, due to limited cooperation of the animal. The reproducibility of the Zeiss SLC system, which offers a direct quality control of the image on the video screen, was tested with 2 animal cataract models, the ultraviolet B (UVB)-induced cataract and the true diabetic cataract in the Brown-Norway rat. 10 rats each of the 2 cataract models and of an untreated control group were photographed in a random order on 3 occasions on the same day by the same photographer. Quality control and densitometry were performed by the same system operator, the measurement window was positioned standardly, coincident with the optical axis of the eye. Statistical comparison was carried out in the capsular layer, that is the area of initial cataract development in both models, and in the cortical layer, into which the diabetic cataract progresses during its later stages of development. The nuclear layer was also evaluated, but has no direct relation to one of the models employed. The results clearly indicate that the reproducibility is predominantly influenced by the homogeneity or inhomogeneity of the cataract model employed. The system operator and the photographer provided that they are well trained, have minor influence on the reproducibility of the Zeiss SLC system.