“Blind focusing”: An objective method for avoiding errors of focus at the television texture analysis system (TAS)

Summary The amount of light energy a particle absorbs does not depend upon correct focus. The change in the path of light rays brought about by defocusing causes absorbing areas to be registered as areas of higher transmittance than when in focus. Already wellknown in photometry, this effect is put to use by the blind focus method at the television texture analysis system (TAS, Leitz). Some chromophores within the object to be measured are compared to a preset value of transmittance, for exampleT=0.40.Only the area representing the structures as dense or denser than the preset density are registered. If the structures are out of focus the size of the registered area is too low, since by defocusing, structures to be measured become pale and diffuse, the correct focus corresponds to the largest area to a preset value of transmittance.     

The influence of preset integer intervals at the densitometer unit of the television texture analysis system (TAS) on DNA quantification

Summary The relative DNA content of Feulgen stained nuclei, or concentrations of stoichiometrically binding dyes can be determined by the television texture analysis system. This determination is based on the measurement of areas at preset integer intervals of transmittance. The method includes an unavoidable error in DNA quantification caused by the smallest preset integer value of transmittance at the densitometer unit. The error at the TAS is calculable by a theoretical reduction of the preset intervals for T. The calculation results in a correcting factor of 04.1% of the whole measured area of a nucleus. The revised relative DNA content can now be calculated by D=·A·1.041.     

Principles and methods for the quantitative determination of feulgen stained DNA with the television texture analysis system (TAS)

Summary The television texture analysis system (TAS, Leitz) has been applied to determination of the relative DNA content of Feulgen stained nuclei. The principles of the method are described and comparative measurements are presented. The determination is based on the measurement of areas at preset intervals of transmittance, taking into consideration the necessary correction of extinction values in relation to area and applying a correction for background. The data obtained can be used for the quantitative determination of DNA, and to provide analytical information for quantitation of structure.Comparison of the mean of extinctions as recorded with the scanning photometric method and those recorded with the television texture analysis method, with which 43 nuclei were investigated in transmittance steps of 0.01, revealed a coefficient of correlation of 0.9938.     

Principles and methods for the quantitative determination of Feulegen stained DNA with the television texture analysis system (TAS).

The television texture analysis system (TAS, Leitz) has been applied to determination of the relative DNA content of Feulgen stained nuclei. The principles of the method are described and comparative measurements are presented. The determination is based on the measurement of areas at preset intervals of transmittance, taking into consideration the necessary correction of extinction values in relation to area and applying a correction for background. The data obtained can be used for the quantitative determination of DNA, and to provide analytical information for quantitation of structure. Comparison of the mean of extinctions as recorded with the scanning photometric method and those recorded with the television texture analysis method, with which 43 nuclei were investigated in transmittance steps of 0.01, revealed a coefficient of correlation of 0.9938.     

The "windows", scales, and bristles of the tropical moth Rothschildia lebeau (Lepidoptera: Saturniidae)

The common Spanish name of the moth Rothschildia lebeau (Saturniidae) is cuatro ventanas (four 'windows'), because it exhibits a transparent oval path in each wing. The scales of the colored areas and the bristles from the "window" were analyzed. We developed a simple device to measure transmittance across the "windows" with an spectrophotometer. A square section of "window" was mounted onto a flat black card and placed onto a clamp that hung in the path of the light - beam of the spectrophotometer. Absorbance was measured at 350 and 550 nm, with the "window" positioned perpendicular to the light beam (incidence of 90 degrees); then the measurements were repeated with the "window" moved at an angle of 45 degrees. Each measurement was replicated 5 times. Wing color spots were analyzed with a light dissection microscope (stereoscope) and with scanning electron microscopy. The scales have a minimum of 4 morphological types, 3 of them showed the typical appearance of unspecialized scales described for other butterflies; whereas the fourth has features particular to this species. On the "window" the scales are transformed in hair-like bristles that do not interfere with light, conferring the transparency that characterizes the "windows". However, if the wing is illuminated at an almost grazing-incidence, they reflect the light as a mirror. Two hypothetical functional explanation for the windows are mimicry and interspecies communication.     

Dynamics of seedling growth acclimation towards altered light conditions can be quantified via GROWSCREEN: a setup and procedure designed for rapid optical phenotyping of different plant species

Using a novel setup, we assessed how fast growth of Nicotiana tabacum seedlings responds to alterations in the light regime and investigated whether starch-free mutants of Arabidopsis thaliana show decreased growth potential at an early developmental stage. Leaf area and relative growth rate were measured based on pictures from a camera automatically placed above an array of 120 seedlings. Detection of total seedling leaf area was performed via global segmentation of colour images for preset thresholds of the parameters hue, saturation and value. Dynamic acclimation of relative growth rate towards altered light conditions occurred within 1 d in N. tabacum exposed to high nutrient availability, but not in plants exposed to low nutrient availability. Increased leaf area was correlated with an increase in shoot fresh and dry weight as well as root growth in N. tabacum. Relative growth rate was shown to be a more appropriate parameter than leaf area for detection of dynamic growth acclimation. Clear differences in leaf growth activity were also observed for A. thaliana. As growth responses are generally most flexible in early developmental stages, the procedure described here is an important step towards standardized protocols for rapid detection of the effects of changes in internal (genetic) and external (environmental) parameters regulating plant growth.     

The combined effects of CO2 concentration and enhanced UV-B radiation on faba bean. 3. Leaf optical properties,pigments, stomatal index and epidermal cell density

Seedlings of Vicia faba L. (cv. Minica) were grown in a factorial experiment in a greenhouse. The purpose of the study was to determine whether CO₂ enrichment and supplemental UV-B radiation affect leaf optical properties and whether the combined effects differ from single factor effects. Seedlings were grown at either 380 mol mol^-1 or 750 mol mol^-1 CO₂ and at four levels of UV-B radiation. After 20 and 40 days of treatment, absorptance, transmittance and reflectance of photosynthetically active radiation (PAR) were measured on the youngest fully developed leaf. On the same leaf, the specific leaf area on a fresh weight basis (SLA_fw), chlorophyll content, UV-B absorbance, transmittance of UV light and stomatal index were measured. UV-B radiation significantly increased PAR absorptance and decreased PAR transmittance. The increased PAR absorptance can be explained by an increased chlorophyll content in response to UV-B radiation. Leaf transmittance of UV radiation decreased with increasing UV-B levels mainly caused by increased absorbance of UV absorbing compounds. UV-B radiation decreased both the stomatal density and epidermal cell density of the abaxial leaf surface, leaving the stomatal index unchanged. Effects of CO₂ enrichment were less pronounced than those of UV-B radiation. The most important CO₂ effect was an increase in stomatal density and epidermal cell density of the adaxial leaf surface. The stomatal index was not affected. No interaction between CO₂ and UV-B radiation was found. The results are discussed in relation to the internal light environment of the leaf.     

Characterisation, prediction and relationships between different wavebands of solar radiation transmitted in the understorey of even-aged oak (Quercus petraea, Q. robur) stands

Solar radiation transmission in forest stands affects many processes, including biomass and diversity of understorey vegetation and tree seedling regeneration (growth and morphogenesis). However, understorey light availability is not easy for forest managers or scientists to measure. Therefore, different models have been developed to predict light transmission in forest stands according to tree or stand structure. However, these models are generally too complex to be used operationally. This paper reports the assessment of light transmission according to stand parameters commonly measured by foresters in inventories. We measured transmittance in 29 even-aged oak stands in France for various wavebands, total solar radiation (TSR, 300–3000 nm), PAR (400–700 nm), red (R, 660 nm) and far-red (FR, 730 nm), and demonstrated that transmittance in a given waveband can be predicted from the measurement of another waveband. The R:FR ratio can be predicted according to TSR or PAR transmittance, but the opposite is also true; PAR or TSR transmittance can be predicted from the R:FR ratio. Transmittance variability was characterised, and the variation coefficient ranged from 5 to 45% with a trend to increase with tree density. By analogy to Beer–Lambert's law, we established that mean daily transmittance for the different wavebands can be assessed according to stand basal area and stand age with good accuracy (R ²>0.74). Results are discussed in comparison with other models based on the principle of parsimony.     

水稻冠层光合有效辐射的时空分布特征

以2个不同株型水稻品种为材料,设置高、中、低3个施氮水平,利用SunScan冠层分析仪于灌浆期系统测定了不同施氮水平下不同株型水稻品种植株形态和冠层内光合有效辐射（PAR）的时空分布状况.结果表明：施氮量对水稻株高、穗弯曲度和茎叶夹角有明显影响;群体叶面积的垂直分布呈中部>上部>下部的分布特征,最大分层叶面积指数（LAI）出现在0.60相对高度处.冠层内平均PAR透光率从顶部向下递减,且在冠层上中部递减迅速,下部递减缓慢;平均PAR透光率随施氮量的增加而递减;平均PAR透光率日变化表现为早晚较低,中午较高;平均PAR透光率随向下累积LAI的增加呈指数递减,群体消光系数K的日变化表现为早晚较高,中午较低,灌浆期的K值介于0.35～0.50.水稻冠层内PAR的三维空间分布表现为冠层上中部水平面上PAR透光率高,光斑面积大;下部水平面上PAR透光率低,光斑少;同一冠层高度水平面上的PAR光强呈不均匀分布.株型紧凑的水稻品种,冠层透光率高,透光率日变化大,群体消光系数小.     

The "intermediate component" of the radiosodium efflux from frog skeletal muscle

The influence of diversity in the size of the cells of the frog's sartorius on the radiosodium efflux from the muscle was investigated. Morphometric analyses of light micrographs of complete cross sections of the muscle were done in the proximal and distal regions. The results were used to predict the shape of the radiosodium washout curve under the following assumptions: the cells differ in size and shape, but each has a single internal pool of exchangeable sodium; the sodium exchange properties of the limiting membranes are the same for all cells; and the diversity of the true areas of the limiting membranes is reflected by the diversity of the apparent areas measured at the light microscopic level. Radiosodium efflux measurements were performed on similar muscles. The model correctly predicted the occurrence of a continuous decline of the fractional loss of radiosodium, which was not due to diffusional delay and which would be interpreted as a second internal compartment in a compartmental analysis, and an effect of short versus long isotope loading intervals on the efflux. It was concluded that the existence of cell size diversity satisfactorily explains the flux data. No "special region" must be postulated.     

"Mind the trap": mindfulness practice reduces cognitive rigidity

Two experiments examined the relation between mindfulness practice and cognitive rigidity by using a variation of the Einstellung water jar task. Participants were required to use three hypothetical jars to obtain a specific amount of water. Initial problems were solvable by the same complex formula, but in later problems ("critical" or "trap" problems) solving was possible by an additional much simpler formula. A rigidity score was compiled through perseverance of the complex formula. In Experiment 1, experienced mindfulness meditators received significantly lower rigidity scores than non-meditators who had registered for their first meditation retreat. Similar results were obtained in randomized controlled Experiment 2 comparing non-meditators who underwent an eight meeting mindfulness program with a waiting list group. The authors conclude that mindfulness meditation reduces cognitive rigidity via the tendency to be "blinded" by experience. Results are discussed in light of the benefits of mindfulness practice regarding a reduced tendency to overlook novel and adaptive ways of responding due to past experience, both in and out of the clinical setting.     

The significance of plant growth-forms as "shelter" for terrestrial animals

Different plant growth-forms such as tussocks and rosettes, were selected as micro ecosystems in an area in which these structures were spatially isolated from each other. These growth forms are important during winter since they serve to regulate temperatures, which is important to invertebrate animals. This was shown with intensive temperature recordings in winter, correlated with numbers of animals. Plant growth-forms are more important in young bare areas where there are fewer plants to provide shelter, than in older areas, where a denser vegetation cover affords more possibilities for shelter. Such differences can be expected to influence the abundance as well as the occurrence of animals during the development of a community.     

不同育秧方式和插植密度下晚籼稻冠层的透光率比较试验

以籼型水稻特三矮为材料,初步研究了水、旱育秧方式和不同基本苗插植密度下生长过程中晚季籼稻冠层的透光率。研究表明:单苗移植的透光率的下降速度高于多苗移植;旱育秧移植的透光率下降速度大于水育秧移植;遮阴生长的其透光率大于不遮阴的。自幼穗分化期起至黄熟期,四种育秧移栽方式的透光率基本相同。     

Producing the cortical phenomenon of "kindling" during acute experiments on cats

Acute experiments on cats enabled spontaneous epileptogenic foci to be produced by means of electrical stimulation of the cerebral cortex. The central area of the suprasylvian gyrus was stimulated by 5-sec stimuli at 3-sec intervals. The strength of neuronal response gradually increased until spontaneous electrical activity set in as stimulation was set at a fixed intensity which had originally evoked local residual discharge at stimulation foci only, i.e., the phenomenon of "kindling" was observed. When "kindling" was produced in the cortical area under study, bursts of spindle activity were recorded on all ECoG with increasing frequency. Recordings of spindle activity then changed to a "spike-wave" pattern of activity. The results of these investigations, which were performed on an isolated strip of cortex, point to the involvement of subcortical structures in mediating "kindling" of the cortical focus.Institute of Clinical and Experimental Neurology, Ministry of Public Health of the Georgian SSR, Tbilisi. Translated from Neirofiziologiya, Vol. 17, No. 5, pp. 601–606, September–October, 1985.     

Comparing Contact Calling Between Black Tufted-Ear Marmosets (<Emphasis Type="Italic">Callithrix penicillata</Emphasis>) in a Noisy Urban Environment and in a Quiet Forest

All habitats have some level of noise but anthropogenic sounds such as those produced by traffic are structurally different from natural sounds, and could cause organisms living in noisy urban areas to modify their vocal communication. We compared temporal and spectral parameters of contact calls in black tufted-ear marmosets (Callithrix penicillata) living in a noisy and a quiet area. From February 2009 to March 2012 we recorded spontaneously produced phee vocalizations by marmosets in two areas in Minas Gerais, Brazil: a noisy urban park (N = 581) in Belo Horizonte, and a quiet natural forest, on Cauaia farm in Matozinhos city (N = 560). We measured the duration, frequencies, and rate of phee vocalizations. We found that marmosets’ phee vocalizations were significantly longer in the noisy area than in the quiet area. The low, high, and dominant frequencies were significantly lower in the noisy area than in the quiet area, and contact calling was less frequent in the noisy area than in the quiet area. We suggest that the differences between marmoset contact calls from noisy and quiet areas are influenced by anthropogenic noise.     

"Schizophrenic" micellization associated with coil-to-helix transitions based on polypeptide hybrid double hydrophilic rod-coil diblock copolymer

A polypeptide hybrid double hydrophilic diblock copolymer (DHBC), poly( N-isopropylacrylamide)- b-poly( l-glutamic acid) (PNIPAM- b-PLGA), was synthesized via the ring-opening polymerization of gamma-benzyl- l-glutamate N-carboxyanhydride (BLG-NCA) using monoamino-terminated PNIPAM as the macroinitiator, followed by deprotection of benzyl groups under alkaline conditions. Containing a thermoresponsive PNIPAM block and a pH-responsive PLGA block, the obtained polypeptide hybrid diblock copolymer molecularly dissolves in aqueous solution at alkaline pH and room temperature but supramolecularly self-assembles into PNIPAM-core micelles at alkaline pH and elevated temperatures and PLGA-core micelles at acidic pH and room temperature accompanied with coil-to-helix transition of the PLGA sequence. The pH- and thermoresponsive "schizophrenic" micellization behavior of PNIPAM- b-PLGA diblock copolymer has been investigated by (1)H NMR, optical transmittance, fluorescence probe measurement, transmission electron microscopy (TEM), dynamic and static laser light scattering (LLS), and circular dichroism (CD) spectroscopy. Moreover, the micellization process was investigated employing stopped-flow light scattering technique. The pH-induced micelle growth of PNIPAM- b-PLGA in aqueous solution exhibits drastically different kinetics compared to that of conventional pH-responsive DHBCs, probably due to the stabilization effects exerted by the formed alpha-helix secondary structures within the PLGA core at low pH. Exhibiting "schizophrenic" micellization, the polypeptide sequence of PNIPAM- b-PLGA can either locate within micelle cores or stabilizing coronas. The incorporation of polypeptide block into DHBCs can endow them with structural versatility, tunable spatial arrangement of chain segments within self-assembled nanostructures, and broader applications in the field of biomedicines.     

Capsule Design for Blue Light Therapy against Helicobacter pylori

A photo-medical capsule that emits blue light for Helicobacter pylori treatment was described in this paper. The system consists of modules for pH sensing and measuring, light-emitting diode driver circuit, radio communication and microcontroller, and power management. The system can differentiate locations by monitoring the pH values of the gastrointestinal tract, and turn on and off the blue light according to the preset range of pH values. Our experimental tests show that the capsule can operate in the effective light therapy mode for more than 32 minutes and the wireless communication module can reliably transmit the measured pH value to a receiver located outside the body.     

Effect of focus on cell detection and recognition by the Cell Analyzer

The effect of defocusing on the quality of signals from live cells detected by an automated image cytometry device, the Cell Analyzer, was examined. The influence of these effects on the ability of this device to automatically locate cells plated into a tissue culture flask was then determined by measuring the performance of cell detection and recognition procedures as a function of focus setting. Acceptable limits for deviation from the optimal focus setting (as determined by microscope objective position) were found to be similar for both these procedures, ranging from 40 microns below to 25 microns above the optimal focus position. These limits were asymmetrical about ideal focus due to a pronounced asymmetry in the effects of positive and negative defocusing on the cell signal.     

Cladocera in shallow lakes from the Ecuadorian Andes show little response to recent climate change

Floating-leaved rhizophytes and pleustophytes are the first barrier to Sun’s rays and significantly affect the light regime of the water column. To evaluate these effects on light attenuation, the reflectance and transmittance spectra variability were examined according to the leaf traits within three plant groups: (1) seed plants with green abaxial surfaces; (2) seed plants with red abaxial surfaces; and (3) ferns with trichomes. Specific leaf area (SLA), chlorophyll a and b, and UV-B and UV-A-absorbing substances differed between these three groups. The ‘spectral signatures’ of floating-leaved seed plants are similar to those of terrestrial seed plants, with a peak in the green region and a pronounced ‘red edge’. Ferns transmitted more light along the whole spectrum compared to other species. Most reflectance and transmittance spectra variability of the first group was explained by SLA. In the second group, 36% of the reflectance spectra variability was explained by tissue density and carotenoids, and 48% of the transmittance spectra variability by carotenoids, anthocyanins and SLA. In ferns, the reflectance spectra variability was mainly explained by chlorophylls, and partly by trichome length and mesophyll thickness, with the transmittance spectra variability significantly affected by chlorophyll b.     

Computerized measurement of the DNA content, areas, and autoradiographic grains of the same nuclei: demonstration that lightly (3H)thymidine-labeled bone marrow cells are predominantly in G0/G1 and G2

A computerized "flying spot" microdensitometer and scanning stage have been combined to measure the cellular DNA content, nuclear areas, and autoradiographic grain areas of the same cells. The slide positions of the Feulgen-stained, (3H)thymidine-labeled cells are mapped with the computerized stage, and nuclear DNA content and areas are then determined by integral absorbance measurements at 588 nm. Following autoradiograph preparation, the cells are relocated and the areas of the autoradiographic grains over each nucleus are measured at a light wavelength (625 nm) and an optical density setting (greater than 0.10) that do not detect the Feulgen stain. The microcomputer calculates the portion of each nucleus covered with autoradiographic grains (grain area proportion, GAP), and it links the GAP value to the DNA content of each nucleus in the computer file for subsequent sorting and analysis. By using this system in a study of mouse bone marrow cells labeled in vivo with (3H)thymidine, we found that all S-phase cells were clearly labeled after 8 or more days of autoradiographic exposure. Prolonged exposures (up to 64 days) led to detection of lightly labeled cells (0.1 less than GAP less than 0.8) with G1/G0 and G2 DNA content.