Análise de imagem em química analítica: empregando metodologias simples e didáticas para entender e prevenir o escurecimento de tecidos vegetais

A simple and didactic experiment was developed for image monitoring of the browning of fruit tissues caused by the enzyme polyphenol oxidase. The procedure, easy and inexpensive, is a valuable tool to teach and demonstrate the redox reaction between the enzyme and the natural polyphenols. To obtain the browning percentage for apple, pear and banana, digital photographs were employed, and the images were analyzed by means of Monte Carlo methods and digital analysis programs. The effects of several experimental conditions were studied, such as pH, light, temperature and the presence of oxygen or anti-oxidants. It was observed that each fruit presented a different condition that better minimized the oxidation process. The absence of oxygen and the application of a bissulphite solution were sufficient to keep the quality of all fruits tested.

Hardcopy quality parameters to ensure structures detection at digital mammography

Objective To develop procedures to ensure consistency of printing quality of digital images, by means of hardcopy quantitative analysis based on a standard image. Materials and Methods Characteristics of mammography DI-ML and general purpose DI-HL films were studied through the QC-Test utilizing different processing techniques in a FujiFilm®-DryPix4000 printer. A software was developed for sensitometric evaluation, generating a digital image including a gray scale and a bar pattern to evaluate contrast and spatial resolution. Results Mammography films showed maximum optical density of 4.11 and general purpose films, 3.22. The digital image was developed with a 33-step wedge scale and a high-contrast bar pattern (1 to 30 lp/cm) for spatial resolution evaluation. Conclusion Mammographic films presented higher values for maximum optical density and contrast resolution as compared with general purpose films. The utilized digital processing technique could only change the image pixels matrix values and did not affect the printing standard. The proposed digital image standard allows greater control of the relationship between pixels values and optical density obtained in the analysis of films quality and printing systems.

Digital surface area assessment of broiler chickens

Surface area (SA) of poultry is an important parameter for heat and mass transfer calculations. Optical approaches, such as the moiré technique (MT), are non-destructive, result in accuracy and speed gains, and preserve the object integrity. The objective of this research was to develop and validate a new protocol for estimating the surface area (SA) of broiler chickens based on the MT. Sixty-six Ross breed broiler chickens (twenty-seven male, thirty-nine female, ages spanning all growth phases) were used in this study. The dimensions (length, width and height) and body mass of randomly selected broiler chickens were evaluated in the laboratory. Chickens were illuminated by a light source, and grids were projected onto the chickens to allow their shape to be determined and recorded. Next, the skin and feathers of the chickens were removed to allow SA to be determined by conventional means. These measurements were then used for calibration and validation. The MT for image analysis was a reliable means of evaluating the three-dimensional shape and SA of broiler chickens. This technique, which is neither invasive nor destructive, is a good alternative to the conventional destructive methods.

Analysis of land use and occupancy in permanent preservation areas according to the hydrography of Ribeirão Água Fria - Bofete, SP - Brazil

The use conflicts are determined by the inadequate occupations of the soil, as it is the case of soil occupation inside of permanent preservation areas. This study aimed to determine the classes of the soil use and if there are conflicts inside of permanent preservation areas along the drainage network of the Água Fria Stream watershed, located in Bofete city - São Paulo, Brazil. It locates geographically between the coordinates: 48°09'30" to 48°18'30" longitude WGr., 22°58'30" to 23°04'30" latitude S, with an area of 15242.84 ha. The map of soil use was elaborated through the interpretation directly in the computer screen of satellite digital image. In the orbital data, the study area is inserted in the quadrant A, of image TM/Landsat - 5, orbit 220, point 76, passage 9/8th/2007. The Geographical Information System used was CartaLinx. The conflict areas of the watershed were obtained from the crossing between the maps of soil use and of PPAs. The results allowed the conclusion that more than half of the area (51.09%) is occupied by pastures, reflex of sandy soils and low fertility. It was also verified that although almost half of the watershed is covered with some type of vegetation (48.78% of natural forest /reforestation), it has approximately a third of permanent preservation areas used inappropriately by pastures (88.15%), reforestation (10.42%) and exposed soil (1.43%), totaling 343.07ha of conflicting areas, in a total of 993.26 ha of PPAs.

X-Ray analysis to assess mechanical damage in sweet corn seeds

The X-ray test is a precise, fast and non-destructive method to detect mechanical damage in seeds. In the present study, the efficiency of X-ray analysis in identifying the extent of mechanical damage in sweet corn seeds and its relationship with germination and vigor was evaluated. Hybrid 'SWB 551' (sh2) seeds with round (R) and flat (F) shapes were classified as large (L), medium (M1, M2 and M3) and small (S), using sieves with round and oblong screens. After artificial exposure to different levels of damage (0, 1, 3, 5 and 7 impacts), seeds were X-rayed (15 kV, 5 min) and submitted to germination (25 °C/5 days) and cold (10 °C/7 days) tests. Digital images of normal and abnormal seedlings and ungerminated seeds from germination and cold tests were jointly analyzed with the seed X-ray images. Results showed that damage affecting the embryonic axis resulted in abnormal seedlings or dead seeds in the germination and cold tests. The X-ray analysis is efficient for identifying mechanical damage in sweet corn seeds, allowing damage severity to be associated with losses in germination and vigor.

Morphometric Analysis of McCoy Cells Inoculated with Cerebrospinal Fluid from Patients with Rabies

To demonstrate the potential of McCoy cells for the isolation of rabies virus from the cerebrospinal (CSF) fluid of a patient with a diagnosis of rabies, McCoy cells were inoculated with CSF from a patient with a clinical diagnosis of rabies and investigated in terms of morphometric aspect using the JAVA analysis system for the quantification of the increased size of infected cells compared to noninfected cells. The cells were also examined in terms of specific staining for the diagnosis of rabies by the method of Sellers for the observation of intracytoplasmic inclusions and by specific immunofluorescence staining for rabies virus. Infected cells showed changes in cell permeability and morphologic modifications which differed significantly compared to normal cells (P<0.001) when analyzed by the Mann-Whitney and Kruskal-Wallis tests. Intense activity of the endoplasmic reticulum was also observed, as indicated by the presence of intracytoplasmic inclusions visualized by specific staining. The present study demonstrated the isolation of rabies virus from the CSF of a patient with rabies, showing that McCoy cells can be used for the laboratory diagnosis of patients suspected to have rabies.

Morphologic and morphometric analysis of Hepatozoon spp. (Apicomplexa, Hepatozoidae) of snakes

Hepatozoon species are the most abundant hemoparasites of snakes. Its identification has been based mainly on the morphologic characterization of the gamonts in the peripheral blood of the vertebrate host and also of the cysts found in the internal organs of the vertebrate and invertebrate hosts. Using a computerized image analysis system, we studied five species of Hepatozoon from recently captured snakes in Botucatu, State of São Paulo, Brazil, to evaluate the importance of the morphology and morphometry of the gamonts for the characterization of Hepatozoon species and to analyze the morphologic changes induced in the erythrocytes by the parasite. The studied species were H. terzii of Boa constrictor amarali, Hepatozoon sp. of Crotalus durissusterrificus, H. philodryasi of Philodryas patagoniensis, and H. migonei and H. cyclagrasi of Hydrodynastes gigas. We observed three different groups, one of them including the species H. terzii, H. philodryasi and Hepatozoon sp. of C. durissus terrificus; and the other two consisting of H. migonei and H. cyclagrasi. Degree of alterations in the erythrocytes was variable and it may be useful for characterization of Hepatozoon species.

Phenotypic plasticity in adult worms of Schistosoma mansoni (Trematoda:Schistosomatidae) evidenced by brightfield and confocal laser scanning microscopies

A comparative morphometric study was performed to identify host-induced morphological alterations in Schistosoma mansoni adult worms. A wild parasite population was obtained from a naturally infected rodent (Nectomys squamipes)and then recovered from laboratory infected C3H/He mice. Furthermore, allopatric worm populations maintained for long-term under laboratory conditions in Swiss Webster mice were passed on to N. squamipes. Suckers and genital system (testicular lobes, uterine egg, and egg spine) were analyzed by a digital system for image analysis. Confocal laser scanning microscopy (CLSM) showed details of the genital system (testicular lobes, vitelline glands, and ovary) and the tegument just below the ventral sucker. Significant morphological changes (p < 0.05) were detected in male worms in all experimental conditions, with no significant variability as assessed by CLSM. Significant changes (p < 0.05) were evident in females from the wild population related to their ovaries and vitelline glands, whereas allopatric females presented differences only in this last character. We conclude that S. mansoni worms present the phenotypic plasticity induced by modifications in the parasite's microenvironment, mainly during the first passage under laboratory conditions.

Development of Schistosoma mansoni worms in mice analyzed by bright field and confocal microscopy

The blood flukes of mammals (Digenea: Schistosomatidae) are among trematodes unique whose adult worms have separeted sexes which are dissimilar in appearance. The developmental features, growth and organogenesis of Schistosoma mansoni were studied in Swiss Webster mice by a digital system for image analysis and confocal microscopy. Data so far obtained showed two phases with significative morphological changes at 3-4 weeks post-infection, and a gradual similar development onwards in the reproductive system and tegument. Our male-dependent phase demonstrated that mating occurs before sexual maturing. At week three, the majority of male worms (59%) had formed the gynaecophoric canal although testicular lobes and tegumental tubercles were absent. By this time, 33% females had an incipient ovary (without cellular differentiation). At week four, 77.2% males presented testicular lobes with few germinative cells while 26% had developing tegumental tubercles. The immature ovary was observed in 69% females. Suckers followed different pattern of growth between male and females. The size of oral and ventral suckers from six-week-old male worms grew abruptly (3.0 fold) more than that of three-week-old. In female worms, maximum growth was attained at week four, reducing in size thereafter. From sixth week onwards, all specimens showed the fully developed reproductive system. Probably, these features are morphological traits which schistosome has experienced from hermaphrodite to dioecy.

Analysis of trophic responses in lesioned brain: focus on basic fibroblast growth factor mechanisms

The actions of fibroblast growth factors (FGFs), particularly the basic form (bFGF), have been described in a large number of cells and include mitogenicity, angiogenicity and wound repair. The present review discusses the presence of the bFGF protein and messenger RNA as well as the presence of the FGF receptor messenger RNA in the rodent brain by means of semiquantitative radioactive in situ hybridization in combination with immunohistochemistry. Chemical and mechanical injuries to the brain trigger a reduction in neurotransmitter synthesis and neuronal death which are accompanied by astroglial reaction. The altered synthesis of bFGF following brain lesions or stimulation was analyzed. Lesions of the central nervous system trigger bFGF gene expression by neurons and/or activated astrocytes, depending on the type of lesion and time post-manipulation. The changes in bFGF messenger RNA are frequently accompanied by a subsequent increase of bFGF immunoreactivity in astrocytes in the lesioned pathway. The reactive astrocytes and injured neurons synthesize increased amount of bFGF, which may act as a paracrine/autocrine factor, protecting neurons from death and also stimulating neuronal plasticity and tissue repair

Effect of age on upper and lower eyelid saccades

To study the effect of age on the metrics of upper and lower eyelid saccades, eyelid movement of two groups of 30 subjects each were measured using computed image analysis. The patients were divided on the basis of age into a younger group (20-30 years) and an older group (60-91 years). Eyelid saccade functions were fitted by the damped harmonic oscillator model. Amplitude and peak velocity were used to compare the effect of age on the saccades of the upper and lower eyelid. There was no statistically significant difference in saccade amplitude between groups for the upper eyelid (mean ± SEM; upward, young = 9.18 ± 0.32 mm, older = 8.93 ± 0.31 mm, t = 0.56, P = 0.58; downward, young = 9.11 ± 0.27 mm, older = 8.86 ± 0.32 mm, t = 0.58, P = 0.56) However, there was a clear decline in the peak velocity of the upper eyelid saccades of older subjects (upward, young = 59.06 ± 2.34 mm/s, older = 50.12 ± 1.95 mm/s, t = 2.93, P = 0.005; downward, young = 71.78 ± 1.78 mm/s, older = 60.29 ± 2.62 mm/s, t = 3.63, P = 0.0006). In contrast, for the lower eyelid there was a clear increase of saccade amplitude in the elderly group (upward, young = 2.27 ± 0.09 mm, older = 2.98 ± 0.15 mm, t = 4.33, P < 0.0001; downward, young = 2.21 ± 0.10 mm, older = 2.96 ± 0.17 mm, t = 3.85, P < 0.001). These data suggest that the aging process affects the metrics of the lid saccades in a different manner according to the eyelid. In the upper eyelid the lower tension exerted by a weak aponeurosis is reflected only on the peak velocity of the saccades. In the lower eyelid, age is accompanied by an increase in saccade amplitude which indicates that the force transmission to the lid is not affected in the elderly.

Radiographic analysis in castor bean seeds (Ricinus communis L.)

Castor bean cropping has great social and economic value, but its production has been affected by factors such as low quality seeds used for sowing. The quick and precise evaluation of seed quality by x-ray test is known as an effective method to evaluate seed lots, but little is known about the interpretation between of the type of radiographic image and the seed quality correlation. The potential of x-ray analysis as a marker of seed physiological quality and as an initial process for the implementation of the use of computer-assisted image analysis was investigated using castor bean seeds of the different cultivars. The seeds were classified according to internal morphology visualized in the radiography and subjected to the germination test, emergency and seedling growth rate. It was possible to identify the different types of internal tissues, morphological and physical damage in castor bean seeds using the x-ray test. Tissues generating translucent images, embryo deformation, or tissues with less than 50% of endosperm reserves or spotted, negatively affected the physiological potential of the seed lots. Radiographic analysis is effective as an instrument to improve castor bean seed lot quality. This non destructive analysis allows the prediction of seedling performance and enabled the selection of high-quality seeds under the standards of a sustainable and precision agriculture

Computerized imaging analysis of seedlings for assessment of physiological potential of wheat seeds

Nowadays, image analysis is one of the most modern tools in evaluating physiological potential of seeds. This study aimed at verifying the efficiency of the seedling imaging analysis to assess physiological potential of wheat seeds. The seeds of wheat, cultivars IAC 370 and IAC 380, each of which represented by five different lots, were stored during four months under natural environmental conditions of temperature (T) and relative humidity (RH), in municipality of Piracicaba, Stated of São Paulo, Brazil. For this, bimonthly assessments were performed to quantify moisture content and physiological potential of seeds by means of tests of: germination, first count, accelerated aging, electrical conductivity, seedling emergence, and computerized analysis of seedlings, using the Seed Vigor Imaging System (SVIS®). It has been concluded that the computerized analyses of seedling through growth indexes and vigor, using the SVIS®, is efficient to assess physiological potential of wheat seeds.