RESUMEN
PURPOSE: The purpose of this study was to evaluate the effects of 16 weeks combined aerobic and resistance training, twice a week, on the physical performance in women treated for gynecological cancer. METHODS: Sixty women (56.9 ± 13.3 years) who had completed curative treatment for gynecological cancer were divided into two groups: a physical training group (PT) (n = 29) or a control group (C) (n = 31). The PT group performed two sessions of combined aerobic and resistance training weekly for 16 weeks. Peak oxygen consumption (VÌO2peak) and one repetition maximum (1RM) of leg press, leg extension, and chest press were measured before group assignment, after 16 weeks and at the 1-year follow-up. RESULTS: A significant increase in VÌO2peak (ml min-1 kg-1) (29.7 ± 8.0 vs. 31.3 ± 8.3, p = .009), leg press (kg) (113.0 ± 27.3 vs. 116.7 ± 29.2, p = .047), leg extension (kg) (44.2 ± 10.1 vs. 48.0 ± 10.6, p < .001), and chest press (kg) (24.5 ± 7.5 vs. 26.9 ± 8.2, p = .001) was seen in the PT group from pre- to post-measurement. The PT group maintained the improved aerobic condition and muscle strength 1 year after the training intervention. In the C group, there were no significant differences between pre- and post-measurements, but a significant decrease (28.2 ± 7.5 vs. 27.0 ± 7.3, p = .040) in the VÌO2peak from post to 1-year follow-up measurements. CONCLUSIONS: Combined aerobic and resistance training twice a week in 16 weeks improves VÌO2peak and maximal strength in women treated for gynecological cancer. The training effects were sustained after 1 year in the PT group.
Asunto(s)
Ejercicio Físico/fisiología , Neoplasias de los Genitales Femeninos/rehabilitación , Entrenamiento de Fuerza/métodos , Femenino , Humanos , Persona de Mediana EdadRESUMEN
We compared the genetic differentiation in the green sea urchin Strongylocentrotus droebachiensis from discrete populations on the NE Atlantic coast. By using eight recently developed microsatellite markers, genetic structure was compared between populations from the Danish Strait in the south to the Barents Sea in the north (56-79°N). Urchins are spread by pelagic larvae and may be transported long distances by northwards-going ocean currents. Two main superimposed patterns were identified. The first showed a subtle but significant genetic differentiation from the southernmost to the northernmost of the studied populations and could be explained by an isolation by distance model. The second pattern included two coastal populations in mid-Norway (65°N), NH and NS, as well as the northernmost population of continental Norway (71°N) FV. They showed a high degree of differentiation from all other populations. The explanation to the second pattern is most likely chaotic genetic patchiness caused by introgression from another species, S. pallidus, into S. droebachiensis resulting from selective pressure. Ongoing sea urchin collapse and kelp forests recovery are observed in the area of NH, NS and FV populations. High gene flow between populations spanning more than 22° in latitude suggests a high risk of new grazing events to occur rapidly in the future if conditions for sea urchins are favourable. On the other hand, the possibility of hybridization in association with collapsing populations may be used as an early warning indicator for monitoring purposes.
RESUMEN
This paper presents a new local volume estimator, the spatial rotator, which is based on measurements on a virtual 3D probe, using computer assisted microscopy. The basic design of the probe builds upon the rotator principle which requires only a few manual intersection markings, thus making the spatial rotator fast to use. Since a 3D probe is involved, it is expected that the spatial rotator will be more efficient than the the nucleator and the planar rotator, which are based on measurements in a single plane. An extensive simulation study shows that the spatial rotator may be more efficient than the traditional local volume estimators. Furthermore, the spatial rotator can be seen as a further development of the Cavalieri estimator, which does not require randomization of sectioning or viewing direction. The tissue may thus be sectioned in any arbitrary direction, making it easy to identify the specific tissue region under study. In order to use the spatial rotator in practice, however, it is necessary to be able to identify intersection points between cell boundaries and test rays in a series of parallel focal planes, also at the peripheral parts of the cell boundaries. In cases where over- and underprojection phenomena are not negligible, they should therefore be corrected for if the spatial rotator is to be applied. If such a correction is not possible, it is needed to avoid these phenomena by using microscopy with increased resolution in the focal plane.
Asunto(s)
Tamaño de la Célula , Células Eucariotas/citología , Procesamiento de Imagen Asistido por Computador/métodos , Microscopía/métodos , Animales , Nucléolo Celular/ultraestructura , Forma de la Célula , Células Eucariotas/ultraestructura , Humanos , Procesamiento de Imagen Asistido por Computador/estadística & datos numéricos , Microscopía/instrumentaciónRESUMEN
INTRODUCTION: The study sought to determine whether mast cell counts in endobronchial biopsies of large airways are related to mast cell counts in the small airways. METHODS: Lungs, obtained postmortem from 10 subjects who had died of non-respiratory causes, were fixed in inflation. Mast cell densities (cells/mm(3)), determined with an optical disector, were compared on histological sections (30 µm thick) of biopsies and small airways stained with human anti-mast cell tryptase. RESULTS: Mean mast cell density over the inner airway wall in biopsies was significantly related to mean mast cell density over the total airway wall in the small airways (r=â¼0.80, p<0.01). A minimum of three biopsies per case was required to demonstrate this relationship. Within relevant count areas, mast cell density was about 1.6-fold higher in the small airways than in the biopsies. CONCLUSIONS: These findings suggest that when mean counts from at least three biopsies per case are used, intersubject comparisons of mast cell density in the inner airway wall in endobronchial biopsies reflect intersubject comparisons of mast cell density over the total airway wall in small airways. This is despite the observation that mast cell densities are generally higher in the small airways.
Asunto(s)
Bronquios/citología , Pulmón/citología , Mastocitos/citología , Adolescente , Adulto , Bronquios/anatomía & histología , Bronquios/inmunología , Recuento de Células , Humanos , Pulmón/inmunología , Masculino , Tamaño de los Órganos , Adulto JovenRESUMEN
The 3D spatial arrangement of particles or cells, for example glial cells, with respect to other particles or cells, for example neurons, can be characterized by the radial number density function, which expresses the number density of so-called 'secondary' particles as a function of their distance to a 'primary' particle. The present paper introduces a new stereological method, the saucor, for estimating the radial number density using thick isotropic uniform random or vertical uniform random sections. In the first estimation step, primary particles are registered in a disector. Subsequently, smaller counting windows are drawn with random orientation around every primary particle, and the positions of all secondary particles within the windows are recorded. The shape of the counting windows is designed such that a large portion of the volume close to the primary particle is examined and a smaller portion of the volume as the distance to the primary object increases. The experimenter can determine the relation between these volumina as a function of the distance by adjusting the parameters of the window graph, and thus reach a good balance between workload and obtained information. Estimation formulae based on the Horvitz-Thompson theorem are derived for both isotropic uniform random and vertical uniform random designs. The method is illustrated with an example where the radial number density of neurons and glial cells around neurons in the human neocortex is estimated using thick vertical sections for light microscopy. The results indicate that the glial cells are clustered around the neurons and the neurons have a tendency towards repulsion from each other.
Asunto(s)
Técnicas Citológicas/métodos , Neocórtex/citología , Neuroglía/citología , Neuronas/citología , Humanos , Microtomía/métodosRESUMEN
We present an unbiased estimator of the total number of alveolar structures distal to the transition from a bronchiole to an alveolar duct system ('ventilatory units', VUs). In species without respiratory bronchioles, including mice, the number of VUs is equivalent to the number of acini. The acinus is a functional unit of gas exchange, defined as a parenchymal unit distal to a terminal bronchiole in which all airways contain alveoli and thus participate in gas exchange. The estimator combines two different estimators of the number of VUs: (1) an estimator derived from the Euler number of all the openings of the bronchial tree and (2) an estimator derived from direct counts of topological changes occurring at bronchiole-alveolar duct junctions. Combining the two estimators eliminates the requirement to be able to identify even vanishingly small pieces of bronchial tissue in physical disectors. We implemented the fractionator estimator in five adult mice lungs using physical fractionators with varying but known sampling fractions (Horvitz-Thompson estimator). We obtained total values of about 4200 VUs (CV = 0.05) in 21-day-old and 4480 (CV = 0.06) in 69-day-old animals. Being fractionator estimates, these total numbers are independent of shrinkage. The densities of VUs per unit volume of tissue (values corrected for tissue shrinkage) were similar in left and right lungs.
Asunto(s)
Biometría/métodos , Pulmón/anatomía & histología , Pulmón/fisiología , Alveolos Pulmonares/anatomía & histología , Alveolos Pulmonares/fisiología , Ventilación Pulmonar , Animales , Femenino , Ratones , Ratones Endogámicos C57BL , Modelos BiológicosRESUMEN
Quantification of tissue properties is improved using the general proportionator sampling and estimation procedure: automatic image analysis and non-uniform sampling with probability proportional to size (PPS). The complete region of interest is partitioned into fields of view, and every field of view is given a weight (the size) proportional to the total amount of requested image analysis features in it. The fields of view sampled with known probabilities proportional to individual weight are the only ones seen by the observer who provides the correct count. Even though the image analysis and feature detection is clearly biased, the estimator is strictly unbiased. The proportionator is compared to the commonly applied sampling technique (systematic uniform random sampling in 2D space or so-called meander sampling) using three biological examples: estimating total number of granule cells in rat cerebellum, total number of orexin positive neurons in transgenic mice brain, and estimating the absolute area and the areal fraction of beta islet cells in dog pancreas. The proportionator was at least eight times more efficient (precision and time combined) than traditional computer controlled sampling.
Asunto(s)
Automatización , Biometría/métodos , Procesamiento de Imagen Asistido por Computador/métodos , Manejo de Especímenes/métodos , Animales , Encéfalo/citología , Recuento de Células , Cerebelo/citología , Perros , Ratones , Ratones Transgénicos , Páncreas/citología , RatasRESUMEN
The proportionator is a novel and radically different approach to sampling with microscopes based on the well-known statistical theory (probability proportional to size-PPS sampling). It uses automatic image analysis, with a large range of options, to assign to every field of view in the section a weight proportional to some characteristic of the structure under study. A typical and very simple example, examined here, is the amount of color characteristic for the structure, marked with a stain with known properties. The color may be specific or not. In the recorded list of weights in all fields, the desired number of fields is sampled automatically with probability proportional to the weight and presented to the expert observer. Using any known stereological probe and estimator, the correct count in these fields leads to a simple, unbiased estimate of the total amount of structure in the sections examined, which in turn leads to any of the known stereological estimates including size distributions and spatial distributions. The unbiasedness is not a function of the assumed relation between the weight and the structure, which is in practice always a biased relation from a stereological (integral geometric) point of view. The efficiency of the proportionator depends, however, directly on this relation to be positive. The sampling and estimation procedure is simulated in sections with characteristics and various kinds of noises in possibly realistic ranges. In all cases examined, the proportionator is 2-15-fold more efficient than the common systematic, uniformly random sampling. The simulations also indicate that the lack of a simple predictor of the coefficient of error (CE) due to field-to-field variation is a more severe problem for uniform sampling strategies than anticipated. Because of its entirely different sampling strategy, based on known but non-uniform sampling probabilities, the proportionator for the first time allows the real CE at the section level to be automatically estimated (not just predicted), unbiased-for all estimators and at no extra cost to the user.
Asunto(s)
Procesamiento de Imagen Asistido por Computador/métodos , Imagenología Tridimensional/métodos , Modelos Estadísticos , Algoritmos , Recuento de Células , Tamaño de la Célula , Simulación por Computador , Técnicas Citológicas/métodos , Microscopía/métodos , Coloración y EtiquetadoRESUMEN
A new stereological probe, the saucer, was used for estimating three-dimensional (3D) spatial distributions of particles around particles. The advantages of the saucer include that the measurements and the results are in 3D and the size and design of the probe enables the investigator to sample a proportion of a suitable size to have a reasonable relationship between workload and the information obtained. In this paper the method is used on vertical sections to investigate the spatial distribution of astrocytes, oligodendrocytes, microglial cells, endothelial cells and secondary neurons around primary neurons in the human neocortex (divided into frontal-, temporal-, parietal- and occipital cortex) of young and old subjects free of neurological or psychological disease to test if age and gender has any influence on the cell distribution in human neocortex. Plots of the spatial distribution of the densities of all cell types did not show any difference between women and men and no difference between brains of young and old subjects. Thus it is concluded that in this small study the spatial distribution of the densities of the different types of cells in brains from individuals free of neurological disorders was independent of age and gender.
Asunto(s)
Envejecimiento/fisiología , Mapeo Encefálico/métodos , Neocórtex/anatomía & histología , Neuroglía/citología , Neuronas/citología , Caracteres Sexuales , Adolescente , Adulto , Anciano de 80 o más Años , Astrocitos/citología , Astrocitos/fisiología , Atrofia/etiología , Atrofia/patología , Atrofia/fisiopatología , Mapeo Encefálico/instrumentación , Recuento de Células/métodos , Células Endoteliales/citología , Células Endoteliales/fisiología , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador/instrumentación , Procesamiento de Imagen Asistido por Computador/métodos , Masculino , Microglía/citología , Microglía/fisiología , Neocórtex/fisiología , Degeneración Nerviosa/etiología , Degeneración Nerviosa/patología , Degeneración Nerviosa/fisiopatología , Neuroglía/fisiología , Neuronas/fisiología , Oligodendroglía/citología , Oligodendroglía/fisiología , Cambios Post MortemRESUMEN
An important step in adoptive immunotherapy in general and specifically with respect to cancer treatment is the initiation of an inflammatory T cell response at the tumor site. Here we suggest a new concept for a controlled inflammatory response in which the intrinsic cytotoxic properties of T cells are upgraded with the properties of nanoparticles transfected into the T cells during the ex vivo expansion process. We report in vitro upgrading of human T cells using PEGylated boron carbide nanoparticles functionalised with a translocation peptide aimed at Boron Neutron Capture Therapy (BNCT). A key finding is that the metabolism of such upgraded human T cells were not affected by a payload of 0.13 pg boron per cell and that the nanoparticles were retained in the cell population after several cell divisions. This is vital for transporting nanoparticles by T cells to the tumor site.
Asunto(s)
Portadores de Fármacos , Inmunoterapia Adoptiva , Nanopartículas/uso terapéutico , Linfocitos T/inmunología , Secuencia de Aminoácidos , Citometría de Flujo , Humanos , Datos de Secuencia MolecularRESUMEN
Stereological estimation of total subcellular quantities in bioscience is presented in this report. Special emphasis is placed on the use of electron microscopy, which under certain circumstances may be combined with light microscopy. Three strategies based on the Cavalieri principle, the disector and local stereological probes through arbitrarily fixed points for estimation of total quantities inside cells are presented. The quantities comprise (total) number, length, surface area, volume or 3D spatial distribution for organelles as well as total amount of gold particles, various compounds or certain cytochemical markers.
Asunto(s)
Recuento de Células/métodos , Microscopía Electrónica , Microtomía , Técnicas Estereotáxicas , Animales , Humanos , Espacio Intracelular/ultraestructura , Modelos Teóricos , Orgánulos/ultraestructuraRESUMEN
The smooth fractionator was introduced in 2002. The combination of a smoothing protocol with a computer-aided stereology tool provides better precision and a lighter workload. This study uses simulation to compare fractionator sampling based on the smooth design, the commonly used systematic uniformly random sampling design and the ordinary simple random sampling design. The smooth protocol is performed using biased information from crude (but fully automatic) image analysis of the fields of view. The different design paradigms are compared using simulation in three different cell distributions with reference to sample size, noise and counting frame position. Regardless of clustering, sample size or noise, the fractionator based on a smooth design is more efficient than the fractionator based on a systematic uniform random design, which is more efficient than a fractionator based on simple random design. The fractionator based on a smooth design is up to four times more efficient than a simple random design.
RESUMEN
The quantification of ultrastructure has been permanently improved by the application of new stereological principles. Both precision and efficiency have been enhanced. Here we report for the first time a fractionator method that can be applied at the electron microscopy level. This new design incorporates a varying sampling fraction paradigm. The method allows for systematic random sampling from blocks of variable slab thickness, thereby eliminating the need for exhaustive serial sectioning through an entire containing space. This novel approach acknowledges the inaccuracy inherent in estimating the total object number using section sampling fractions based on the average thickness of sections of variable thicknesses. As an alternative, this approach estimates the correct particle section sampling probability based on an estimator of the Horvitz-Thompson type, resulting in a theoretically more satisfying and accurate estimate of the expected number of particles for the defined containing space.
Asunto(s)
Hipocampo/ultraestructura , Microscopía Electrónica/métodos , Sinapsis , Animales , Hipocampo/citología , Ratones , Probabilidad , Técnicas EstereotáxicasRESUMEN
In this paper we present surface modification strategies of boron carbide nanoparticles, which allow for bioconjugation of the transacting transcriptional activator (TAT) peptide and fluorescent dyes. Coated nanoparticles can be translocated into murine EL4 thymoma cells and B16 F10 malignant melanoma cells in amounts as high as 0.3 wt. % and 1 wt. %, respectively. Neutron irradiation of a test system consisting of untreated B16 cells mixed with B16 cells loaded with boron carbide nanoparticles were found to inhibit the proliferative capacity of untreated cells, showing that cells loaded with boron-containing nanoparticles can hinder the growth of neighboring cells upon neutron irradiation. This could provide the first step toward a T cell-guided boron neutron capture therapy.
Asunto(s)
Compuestos de Boro , Terapia por Captura de Neutrón de Boro , Nanoestructuras/química , Linfocitos T/metabolismo , Animales , Compuestos de Boro/química , Compuestos de Boro/metabolismo , Línea Celular Tumoral/efectos de la radiación , Proliferación Celular , Melanoma , Ratones , Estructura Molecular , Tamaño de la PartículaRESUMEN
In a mouse model, we demonstrate how to obtain a direct, unbiased estimate of the total number of adoptively transferred cells in a variety of organs at different time points. The estimate is obtained by a straightforward method based on the optical fractionator principle. Specifically, non-stimulated C57BL/6J mouse splenocytes were labelled with carboxyfluorescein diacetate succinimidyl ester (CFSE) and adoptively transferred to normal C57BL/6J mice by intravenous injection. The total number of CFSE-positive cells was subsequently determined in lung, spleen, liver, kidney, and inguinal lymph node at six different time points following adoptive transfer (from 60 s to 1 week), providing a quantitative estimate of the organ distribution of the transferred cells over time. These estimates were obtained by microscopy of uniform samples of thick sections from the respective organs. Importantly, the samples were chosen and prepared in accordance with the optical fractionator principle. We demonstrate that the method is simple, precise, and well suited for quantitative immunological studies.
Asunto(s)
Traslado Adoptivo , Técnicas Inmunológicas , Animales , Biometría , Recuento de Células , Separación Celular , Femenino , Fluoresceínas , Colorantes Fluorescentes , Ratones , Ratones Endogámicos C57BL , Microscopía Fluorescente , Especificidad de Órganos , Bazo/citología , Succinimidas , Factores de TiempoRESUMEN
Boron carbide nanoparticles are proposed as a system for T cell-guided boron neutron capture therapy. Nanoparticles were produced by ball milling in various atmospheres of commercially available boron carbide. The physical and chemical properties of the particles were investigated using transmission electron microscopy, photon correlation spectroscopy, X-ray photoelectron spectroscopy, X-ray diffraction, vibrational spectroscopy, gel electrophoresis and chemical assays and reveal profound changes in surface chemistry and structural characteristics. In vitro thermal neutron irradiation of B16 melanoma cells incubated with sub-100 nm nanoparticles (381.5 microg/g (10)B) induces complete cell death. The nanoparticles alone induce no toxicity.
Asunto(s)
Compuestos de Boro/síntesis química , Compuestos de Boro/uso terapéutico , Terapia por Captura de Neutrón de Boro/métodos , Melanoma/patología , Melanoma/radioterapia , Nanoestructuras/química , Radioinmunoterapia/métodos , Linfocitos T/trasplante , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de la radiación , Marcaje Isotópico , Ratones , Tamaño de la Partícula , Radioisótopos/química , Radioisótopos/uso terapéutico , Radiofármacos/síntesis química , Resultado del TratamientoRESUMEN
The dentate nucleus is phylogenetically the most recent nucleus in the cerebellum. Owing to its connections to the thalamus and the prefrontal cortex it may be involved in the symptomathology in schizophrenia and other psychiatric illnesses. In this stereological study we implemented the smooth fractionator, which combines the unbiased principles of the optical fractionator with a new and more efficient sampling strategy to the dentate nucleus. The smooth fractionator represents the most efficient sampling strategy described so far in stereology, in terms of reducing the sampling variance and thus increasing the efficiency. It is the first application of the smooth fractionator to human brain tissue and presents estimations of total number of neurons in the dentate nuclei of eight patients with schizophrenia compared to eight control persons. The total number of neurons in the dentate nucleus was estimated to 3.36 x 10(6) in subjects with schizophrenia, which was not statistically significant different from 3.65 x 10(6) in control subjects (P = 0.63). The advantages and disadvantages of the smooth fractionator method are discussed and its precision in practical application is estimated.
Asunto(s)
Núcleos Cerebelosos/patología , Neuronas/patología , Esquizofrenia/patología , Adulto , Anciano , Estudios de Casos y Controles , Recuento de Células , Procesamiento Automatizado de Datos , Humanos , Procesamiento de Imagen Asistido por Computador , Masculino , Persona de Mediana EdadRESUMEN
We describe a new estimator of the volume of axially convex objects from total vertical projections with known position of the vertical axis. The estimator combines the Cavalieri method with the known formula for area in terms of the support function of a convex body. We examine the accuracy of the proposed estimator for ellipsoidal objects having exactly known support function and volume. In addition, we illustrate practical problems of accuracy by implementing the method for some biological products.
Asunto(s)
Microscopía/métodos , Biología/métodos , Modelos TeóricosRESUMEN
Estimation of alveolar number in the lung has traditionally been done by assuming a geometric shape and counting alveolar profiles in single, independent sections. In this study, we used the unbiased disector principle to estimate the Euler characteristic (and thereby the number) of alveolar openings in rat lungs and rhesus monkey lung lobes and to obtain robust estimates of average alveolar volume. The estimator of total alveolar number was based on systematic, uniformly random sampling using the fractionator sampling design. The number of alveoli in the rat lung ranged from 17.3 x 10(6) to 24.6 x 10(6), with a mean of 20.1 x 10(6). The average number of alveoli in the two left lung lobes in the monkey ranged from 48.8 x 10(6) to 67.1 x 10(6) with a mean of 57.7 x 10(6). The coefficient of error due to stereological sampling was of the order of 0.06 in both rats and monkeys and the biological variation (coefficient of variance between individuals) was 0.15 in rat and 0.13 in monkey (left lobe, only). Between subdivisions (left/right in rat and cranial/caudal in monkey) there was an increase in variation, most markedly in the rat. With age (2-13 years) the alveolar volume increased 3-fold (as did parenchymal volume) in monkeys, but the alveolar number was unchanged. This study illustrates that use of the Euler characteristic and fractionator sampling is a robust and efficient, unbiased principle for the estimation of total alveolar number in the lung or in well-defined parts of it.
Asunto(s)
Fraccionamiento Celular/métodos , Alveolos Pulmonares/citología , Factores de Edad , Animales , Recuento de Células/instrumentación , Recuento de Células/métodos , Fraccionamiento Celular/instrumentación , Tamaño de la Célula/fisiología , Femenino , Macaca mulatta , Masculino , Alveolos Pulmonares/fisiología , Ratas , Ratas Wistar , Estadística como AsuntoRESUMEN
A modification of the general fractionator sampling technique called the smooth fractionator is presented. It may be used in almost every situation in which sampling is performed from distinct items that are uniquely defined, often they are physically separated items or clusters like pieces, blocks, slabs, sections, etc. To each item is associated a 'guesstimate' or an associated variable with a more-or-less close--and possibly biased--relationship to the content of the item. The smooth fractionator is systematic sampling among the items arranged according to the guesstimates in a unique, symmetric sequence with one peak and minimal jumps. The smooth fractionator is both very simple to implement and so efficient that it should probably always be used unless the natural sequence of the sampling items is equally smooth. So far, there is no theory for the prediction of the efficiency of smooth fractionator designs in general, and their properties are therefore illustrated with a range of real and simulated examples. At the cost of a slightly more elaborate sampling scheme, it is, however, always possible to obtain an unbiased estimate of the real precision and of some of the variance components. The only real practical problem for always obtaining a high precision with the smooth fractionator is specimen inhomogeneity, but that is detectable at almost no extra cost. With careful designs and for sample sizes of about 10, the sampling variation for the primary, smooth fractionator sampling step may in practice often be small enough to be ignored.