Complementarity of radioautographic and immunohistochemical techniques for localizing neuroreceptors at the light and electron microscopy level

To assess relationships between neuropeptide-binding sites and receptor proteins in rat brain, the distribution of radioautographically labeled somatostatin and neurotensin-binding sites was compared to that of immunolabeled sst2A and NTRH receptor subtypes, respectively. By light microscopy, immunoreactive sst2A receptors were either confined to neuronal perikarya and dendrites or diffusely distributed in tissue. By electron microscopy, areas expressing somatodendritic sst2A receptors displayed only low proportions of membrane-associated, as compared to intracellular, receptors. Conversely, regions displaying diffuse sst2A labeling exhibited higher proportions of membrane-associated than intracellular receptors. Furthermore, the former showed only low levels of radioautographically labeled somatostatin-binding sites whereas the latter contained high densities of somatostatin-binding suggesting that membrane-associated receptors are preferentially recognized by the radioligand. In the case of NTRH receptors, there was a close correspondence between the light microscopic distribution of NTRH immunoreactivity and that of labeled neurotensin-binding sites. Within the substantia nigra, the bulk of immuno- and autoradiographically labeled receptors were associated with the cell bodies and dendrites of presumptive DA neurons. By electron microscopy, both markers were detected inside as well as on the surface of labeled neurons. At the level of the plasma membrane, their distribution was highly correlated and characterized by a lack of enrichment at the level of synaptic junctions and by a homogeneous distribution along the remaining neuronal surface, in conformity with the hypothesis of an extra-synaptic action of this neuropeptide. Inside labeled dendrites, there was a proportionally higher content of immunoreactive than radiolabeled receptors. Some of the immunolabeled receptors not recognized by the radioligand were found in endosome-like organelles suggesting that, as in the case of sst2A receptors, they may have undergone endocytosis subsequent to binding to the endogenous peptide

Acute promyelocytic leukemia: the study of t(15;17) translocation by fluorescent in situ hybridization, reverse transcriptase-polymerase chain reaction and cytogenetic techniques

Acute promyelocytic leukemia (AML M3) is a well-defined subtype of leukemia with specific and peculiar characteristics. Immediate identification of t(15;17) or the PML/RARA gene rearrangement is fundamental for treatment. The objective of the present study was to compare fluorescent in situ hybridization (FISH), reverse transcriptase-polymerase chain reaction (RT-PCR) and karyotyping in 18 samples (12 at diagnosis and 6 after treatment) from 13 AML M3 patients. Bone marrow samples were submitted to karyotype G-banding, FISH and RT-PCR. At diagnosis, cytogenetics was successful in 10 of 12 samples, 8 with t(15;17) and 2 without. FISH was positive in 11/12 cases (one had no cells for analysis) and positivity varied from 25 to 93% (mean: 56%). RT-PCR was done in 6/12 cases and all were positive. Four of 8 patients with t(15;17) presented positive RT-PCR as well as 2 without metaphases. The lack of RT-PCR results in the other samples was due to poor quality RNA. When the three tests were compared at diagnosis, karyotyping presented the translocation in 80% of the tested samples while FISH and RT-PCR showed the PML/RARA rearrangement in 100% of them. Of 6 samples evaluated after treatment, 3 showed a normal karyotype, 1 persistence of an abnormal clone and 2 no metaphases. FISH was negative in 4 samples studied and 2 had no material for analysis. RT-PCR was positive in 4 (2 of which showed negative FISH, indicating residual disease) and negative in 2. When the three tests were compared after treatment, they showed concordance in 2 of 6 samples or, when there were not enough cells for all tests, concordance between karyotype and RT-PCR in one. At remission, RT-PCR was the most sensitive test in detecting residual disease, as expected (positive in 4/6 samples). An incidence of about 40% of 5' breaks and 60% of 3' breaks, i.e., bcr3 and bcr1/bcr2, respectively, was observed.

A conceptual and practical overview of cDNA microarray technology: implications for basic and clinical sciences

cDNA microarray is an innovative technology that facilitates the analysis of the expression of thousands of genes simultaneously. The utilization of this methodology, which is rapidly evolving, requires a combination of expertise from the biological, mathematical and statistical sciences. In this review, we attempt to provide an overview of the principles of cDNA microarray technology, the practical concerns of the analytical processing of the data obtained, the correlation of this methodology with other data analysis methods such as immunohistochemistry in tissue microarrays, and the cDNA microarray application in distinct areas of the basic and clinical sciences.

Management zones in agriculture acording to the soil and landscape variables

The study of spatial variability of soil and plants attributes, or precision agriculture, a technique that aims the rational use of natural resources, is expanding commercially in Brazil. Nevertheless, there is a lack of mathematical analysis that supports the correlation of these independent variables and their interactions with the productivity, identifying scientific standards technologically applicable. The aim of this study was to identify patterns of soil variability according to the eleven physical and seven chemical indicators in an agricultural area. It was used two multivariate techniques: the hierarchical cluster analysis (HCA) and the principal component analysis (PCA). According to the HCA, the area was divided into five management zones: zone 1 with 2.87ha, zone 2 with 0.8ha, zone 3 with 1.84ha, zone 4 with 1.33ha and zone 5 with 2.76ha. By the PCA, it was identified the most important variables within each zone: V% for the zone 1, CTC in the zone 2, levels of H+Al in the zone 4 and sand content and altitude in the zone 5. The zone 3 was classified as an intermediate zone with characteristics of all others. According to the results it is concluded that it is possible to separate into groups (management zones) samples with the same patterns of variability by the multivariate statistical techniques.

Distinction of males of the Lutzomyia intermedia (Lutz & Neiva, 1912) species complex by ratios between dimensions and by an artificial neural network (Diptera: Psychodidae, Phlebotominae)

The females of the two species of the Lutzomyia intermedia complex can be easily distinguished, but the males of each species are quite similar. The ratios between the extra-genital and the genital structures of L. neivai are larger than those of L. intermedia s. s., according to ANOVA. An artificial neural network was trained with a set of 300 examples, randomly taken from a sample of 358 individuals. The input vectors consisted of several ratios between some structures of each insect. The model was tested on the remaining 58 insects, 56 of which (96.6%) were correctly identified. This ratio of success can be considered remarkable if one takes into account the difficulty of attaining comparable results using traditional statistical techniques.

Uma introdução à análise exploratória de dados multivariados

The modern technological ability to handle large amounts of information confronts the chemist with the necessity to re-evaluate the statistical tools he routinely uses. Multivariate statistics furnishes theoretical bases for analyzing systems involving large numbers of variables. The mathematical calculations required for these systems are no longer an obstacle due to the existence of statistical packages that furnish multivariate analysis options. Here basic concepts of two multivariate statistical techniques, principal component and hierarchical cluster analysis that have received broad acceptance for treating chemical data are discussed.

Determination of cetirizine in tablets and compounded capsules: comparative study between CE and HPLC

A capillary electrophoresis (CE) method was developed and validated for determination of cetirizine dihydrochloride in tablets and compounded capsules. The electrophoretic separation was performed in an uncoated fused-silica capillary (40 cm x 50 μm i.d.) using 20 mmol L-1 sodium tetraborate buffer (pH 9.3) as background electrolyte, a hydrodinamic sample injection at 50 mBar for 5 s, 20 KV applied voltage at 25 °C, and detection at 232 nm. The proposed method was compared with the high performance liquid chromatographic (HPLC) method previously validated for this drug, and statistical analysis showed no significant difference between the techniques.

Authenticity study of Phyllanthus species by NMR and FT-IR Techniques coupled with chemometric methods

The importance of medicinal plants and their use in industrial applications is increasing worldwide, especially in Brazil. Phyllanthus species, popularly known as "quebra-pedras" in Brazil, are used in folk medicine for treating urinary infections and renal calculus. This paper reports an authenticity study, based on herbal drugs from Phyllanthus species, involving commercial and authentic samples using spectroscopic techniques: FT-IR, ¹H HR-MAS NMR and ¹H NMR in solution, combined with chemometric analysis. The spectroscopic techniques evaluated, coupled with chemometric methods, have great potential in the investigation of complex matrices. Furthermore, several metabolites were identified by the NMR techniques.

Techniques of radioautography for medical and biological research

Standard techniques for radioautography used in biological and medical research can be classified into three categories, i.e., macroscopic radioautography, light microscopic radioautography and electron microscopic radioautography. The routine techniques used in these three procedures are described. With regard to macroscopic radioautography, whole body radioautography is a standard technique which employs freezing and cryosectioning and can demonstrate organ distributions of both soluble and insoluble compounds. In contrast, in light and electron microscopic radioautography, soluble and insoluble techniques are separated. In order to demonstrate insoluble labeled compounds, conventional chemical fixations such as formalin for light microscopy or buffered glutaraldehyde and osmium tetroxide for both light and electron microscopy followed by dehydration, embedding and wet-mounting applications of radioautographic emulsions can be used. For the demonstration of soluble labeled compounds, however, cryotechniques such as cryofixation, cryosectioning, freeze-drying, freeze-substitution followed by dry-sectioning and dry-mounting radioautography should be employed both for light and electron microscopy. The outlines of these techniques, which should be utilized in various fields of biological and medical research, are described in detail

Statistical analysis of event-related potential elicited by verb-complement merge in Brazilian Portuguese

An interesting fact about language cognition is that stimulation involving incongruence in the merge operation between verb and complement has often been related to a negative event-related potential (ERP) of augmented amplitude and latency of ca. 400 ms - the N400. Using an automatic ERP latency and amplitude estimator to facilitate the recognition of waves with a low signal-to-noise ratio, the objective of the present study was to study the N400 statistically in 24 volunteers. Stimulation consisted of 80 experimental sentences (40 congruous and 40 incongruous), generated in Brazilian Portuguese, involving two distinct local verb-argument combinations (nominal object and pronominal object series). For each volunteer, the EEG was simultaneously acquired at 20 derivations, topographically localized according to the 10-20 International System. A computerized routine for automatic N400-peak marking (based on the ascendant zero-cross of the first waveform derivative) was applied to the estimated individual ERP waveform for congruous and incongruous sentences in both series for all ERP topographic derivations. Peak-to-peak N400 amplitude was significantly augmented (P < 0.05; one-sided Wilcoxon signed-rank test) due to incongruence in derivations F3, T3, C3, Cz, T5, P3, Pz, and P4 for nominal object series and in P3, Pz and P4 for pronominal object series. The results also indicated high inter-individual variability in ERP waveforms, suggesting that the usual procedure of grand averaging might not be considered a generally adequate approach. Hence, signal processing statistical techniques should be applied in neurolinguistic ERP studies allowing waveform analysis with low signal-to-noise ratio.

STATISTICAL ANALYSYS OF THE SCFE OF A BRAZILAN MINERAL COAL

The influence of some process variables on the productivity of the fractions (liquid yield times fraction percent) obtained from SCFE of a Brazilian mineral coal using isopropanol and ethanol as primary solvents is analyzed using statistical techniques. A full factorial 23 experimental design was adopted to investigate the effects of process variables (temperature, pressure and cosolvent concentration) on the extraction products. The extracts were analyzed by the Preparative Liquid Chromatography-8 fractions method (PLC-8), a reliable, non destructive solvent fractionation method, especially developed for coal-derived liquids. Empirical statistical modeling was carried out in order to reproduce the experimental data. Correlations obtained were always greater than 0.98. Four specific process criteria were used to allow process optimization. Results obtained show that it is not possible to maximize both extract productivity and purity (through the minimization of heavy fraction content) simultaneously by manipulating the mentioned process variables.

Characterization of commercial cooked hams according to physicochemical, sensory, and textural parameters using chemometrics

Cooked ham is considered a high-value product due to the quality of its raw material. Although its consumption is still low in Brazil, it is increasing due to the rising purchasing power of sectors of the population. This study aimed to assess the microbiological, physicochemical, rheological, and sensory quality of cooked hams (n=11) marketed in Brazil. All samples showed microbiological results within the standards established by Brazilian legislation. Eight of the eleven samples studied met all the legal requirements; two samples violated the standards due to the addition of starch; one sample had lower protein content than the minimum required, and another one had sodium content higher than that stated on the label. The use of Hierarchical Cluster Analysis allowed the agglomeration of the samples into three groups with distinct quality traits and with significant differences in moisture content, chromaticity, syneresis, and heating and freezing loss. Principal Component Analysis showed that the samples which correlated to higher sensory acceptance regarding flavor and overall acceptability were those with higher moisture, protein, fat, and luminosity values. This study confirmed the efficacy of multivariate statistical techniques in assessing the quality of commercial cooked hams and in indicating the physicochemical parameters associated with the perception of product quality.

Health promoting and sensory properties of phenolic compounds in food

Phenolic compounds have been extensively studied in recent years. The presence of these compounds in various foods has been associated with sensory and health promoting properties. These products from the secondary metabolism of plants act as defense mechanisms against environmental stress and attack by other organisms. They are divided into different classes according to their chemical structures. The objective of this study was to describe the different classes of phenolic compounds, the main food sources and factors of variation, besides methods for the identification and quantification commonly used to analyze these compounds. Moreover, the role of phenolic compounds in scavenging oxidative stress and the techniques of in vitro antioxidant evaluation are discussed. In vivo studies to evaluate the biological effects of these compounds and their impact on chronic disease prevention are presented as well. Finally, it was discussed the role of these compounds on the sensory quality of foods.

BRAND EQUITY OF LAHORE FORT AS A TOURISM DESTINATION BRAND

ABSTRACTStudies that measure the brand equity of destination brands by using the Customer-Based Brand Equity (CBBE) model in a developing country context are scarce. The present study investigates the destination brand equity of the Lahore Fort by employing the CBBE model in a developing country context of Pakistan. Following the positivist tradition, we adopted a survey-based approach to collect data from 237 tourists visiting the Lahore Fort. Data were collected through a questionnaire developed to explain the relationship of brand awareness, brand image, brand association, and brand loyalty with Lahore Fort’s overall brand equity. We used various robust statistical techniques such as correlation, regression and confirmatory factor analysis (using PLS method) to reach meaningful conclusions and found that brand image and brand associations positively contribute to brand loyalty. Furthermore, brand loyalty significantly contributes towards overall brand equity. Pragmatically, this study measures the customer based brand equity of the Lahore Fort, a destination brand. The results are useful as they suggest a few strategies that can help policy makers to enhance Lahore Fort’s brand performance.

Handling random errors and biases in methods used for short-term dietary assessment

Epidemiological studies have shown the effect of diet on the incidence of chronic diseases; however, proper planning, designing, and statistical modeling are necessary to obtain precise and accurate food consumption data. Evaluation methods used for short-term assessment of food consumption of a population, such as tracking of food intake over 24h or food diaries, can be affected by random errors or biases inherent to the method. Statistical modeling is used to handle random errors, whereas proper designing and sampling are essential for controlling biases. The present study aimed to analyze potential biases and random errors and determine how they affect the results. We also aimed to identify ways to prevent them and/or to use statistical approaches in epidemiological studies involving dietary assessments.