Effects of heat stress on development, quality and survival of Bos indicus and Bos taurus embryos produced in vitro

Heat stress is an important cause of poor development and low survival rates in bovine embryos. Experiments were conducted to test the hypothesis that Bos indicus embryos are more resistant to heat stress than are Bos taurus embryos. In experiment 1, Nelore and Jersey embryos from oocyte pick-up-derived oocytes were submitted to heat stress (96 hours post-insemination, 41 °C, 6 hours), developmental ratios were assessed at Day 7 (Day 0 = day of fertilization), and blastocysts were frozen for RNA extraction. Experiment 2 evaluated expression of COX2, CDX2, HSF1, and PLAC8 in previously frozen blastocysts. In experiment 3, Nellore and Angus embryos from oocyte pick-up-derived oocytes were submitted to heat stress (96 hours post-insemination, 41 °C, 12 hours) and transferred to recipients on Day 7. In experiment 4, embryos developed as in experiment 3 were fixed for Terminal deoxynucleotidyl transferase dUTP nick end labeling labeling and total cell counting. In experiment 1, heat stress decreased the percentage of Jersey oocytes that became blastocysts, but had no effect on Nellore embryos (34.6%, 25.0%, 39.5%, and 33.0% for Jersey control, Jersey heat-stressed, Nellore control, and Nellore heat-stressed oocytes, respectively; P < 0.05). In experiment 2, heat stress decreased (P < 0.05) expression of CDX2 and PLAC8, with higher expression of these genes in Nellore embryos than in Jersey embryos. Heat stress also decreased (P < 0.05) expression of COX2 in Jersey embryos, but had no effect on Nellore embryos. Expression of HSF1 was decreased (P < 0.05) by heat stress in both breeds, with a greater effect in Nellore embryos. In experiment 3, heat stress tended (P = 0.1) to decrease the percentage of pregnancies among cows (Day 30 to 35) that received Angus embryos. In experiment 4, heat stress increased (P < 0.05) the percentage of apoptotic blastomeres, but had no breed-specific effects. In addition, Nellore embryos had fewer (P < 0.05) Terminal deoxynucleotidyl transferase dUTP nick end labeling- positive blastomeres than did Angus embryos. We concluded that the detrimental effects of heat stress were dependent upon embryo breed and were more evident in Bos taurus embryos than in Bos indicus embryos. © 2013 Elsevier Inc.

Macroeconomy for development: countercyclical policies and production sector transformation

Includes bibliography

Development and evaluation of praziquantel solid dispersions in sodium starch glycolate

Purpose: To develop and characterize solid dispersions of praziquantel (PZQ) with sodium starch glycolate (SSG) for enhanced drug solubility. Methods: PZQ solid dispersion (SD) was prepared using co-precipitation method by solvent evaporation. The ratios of PZQ to SSG were 2:1, 1:1, 1:2, 1:3 (w/w). PZQ solubility was evaluated in purified water, and PZQ dissolution test was carried out in 0.1N HCl. Structural characterization of the dispersions was accomplished by x-ray diffraction (XRD) and infrared spectroscopy (FTIR) while the external morphology of the SDs, SSG and PZQ were studied by scanning electron microscopy (SEM). Mucoadhesion properties of the SD (1:3) and SSG, on mucin disks were examined using texture profile analysis. Results: The highest solubility was obtained with 1:3 solid dispersion, with PZQ solubility of 97.31 %, which is 3.65-fold greater than the solubility of pure PZQ and physical misture (PM, 1:3). XRD results indicate a reduction in PZQ crystallinity while infrared spectra showed that the functional groups of PZQ and SSG were preserved. SEM showed that the physical structure of PZQ was modified from crystalline to amorphous. The amount of PZQ in PM and SD (1:3) that dissolved in 60 min was 70 and 88 %, respectively, and these values increased to 76 and 96 %, respectively. The solid dispersion reduced the mucoadhesive property of the glycolate. Conclusion: Solid dispersion formulation using SSG is a good alternative approach for increasing the dissolution rate of PZQ. © Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, 300001 Nigeria. All rights reserved.

Development and validation of a D-loop mtDNA SNP assay for the screening of specimens in forensic casework

Mitochondrial DNA (mtDNA) analysis is usually a last resort in routine forensic DNA casework. However, it has become a powerful tool for the analysis of highly degraded samples or samples containing too little or no nuclear DNA, such as old bones and hair shafts. The gold standard methodology still constitutes the direct sequencing of polymerase chain reaction (PCR) products or cloned amplicons from the HVS-1 and HVS-2 (hypervariable segment) control region segments. Identifications using mtDNA are time consuming, expensive and can be very complex, depending on the amount and nature of the material being tested. The main goal of this work is to develop a less labour-intensive and less expensive screening method for mtDNA analysis, in order to aid in the exclusion of non-matching samples and as a presumptive test prior to final confirmatory DNA sequencing. We have selected 14 highly discriminatory single nucleotide polymorphisms (SNPs) based on simulations performed by Salas and Amigo (2010) [1] to be typed using SNaPShotTM (Applied Biosystems, Foster City, CA, USA). The assay was validated by typing more than 100 HVS-1/HVS-2 sequenced samples. No differences were observed between the SNP typing and DNA sequencing when results were compared, with the exception of allelic dropouts observed in a few haplotypes. Haplotype diversity simulations were performed using 172 mtDNA sequences representative of the Brazilian population and a score of 0.9794 was obtained when the 14 SNPs were used, showing that the theoretical prediction approach for the selection of highly discriminatory SNPs suggested by Salas and Amigo (2010) [1] was confirmed in the population studied. As the main goal of the work is to develop a screening assay to skip the sequencing of all samples in a particular case, a pair-wise comparison of the sequences was done using the selected SNPs. When both HVS-1/HVS-2 SNPs were used for simulations, at least two differences were observed in 93.2% of the comparisons performed. The assay was validated with casework samples. Results show that the method is straightforward and can be used for exclusionary purposes, saving time and laboratory resources. The assay confirms the theoretic prediction suggested by Salas and Amigo (2010) [1]. All forensic advantages, such as high sensitivity and power of discrimination, as also the disadvantages, such as the occurrence of allele dropouts, are discussed throughout the article. © 2013 Elsevier B.V.

The Development of a Universal In Silico Predictor of Protein-Protein Interactions

Protein-protein interactions (PPIs) are essential for understanding the function of biological systems and have been characterized using a vast array of experimental techniques. These techniques detect only a small proportion of all PPIs and are labor intensive and time consuming. Therefore, the development of computational methods capable of predicting PPIs accelerates the pace of discovery of new interactions. This paper reports a machine learning-based prediction model, the Universal In Silico Predictor of Protein-Protein Interactions (UNISPPI), which is a decision tree model that can reliably predict PPIs for all species (including proteins from parasite-host associations) using only 20 combinations of amino acids frequencies from interacting and non-interacting proteins as learning features. UNISPPI was able to correctly classify 79.4% and 72.6% of experimentally supported interactions and non-interacting protein pairs, respectively, from an independent test set. Moreover, UNISPPI suggests that the frequencies of the amino acids asparagine, cysteine and isoleucine are important features for distinguishing between interacting and non-interacting protein pairs. We envisage that UNISPPI can be a useful tool for prioritizing interactions for experimental validation. © 2013 Valente et al.

Noradrenergic neurotransmission within the bed nucleus of the stria terminalis modulates the retention of immobility in the rat forced swimming test

The bed nucleus of the stria terminalis (BNST) is a limbic structure that has a direct influence on the autonomic, neuroendocrine, and behavioral responses to stress. It was recently reported that reversible inactivation of synaptic transmission within this structure causes antidepressant-like effects, indicating that activation of the BNST during stressful situations would facilitate the development of behavioral changes related to the neurobiology of depression. Moreover, noradrenergic neurotransmission is abundant in the BNST and has an important role in the regulation of emotional processes related to the stress response. Thus, this study aimed to test the hypothesis that activation of adrenoceptors within the BNST facilitates the development of behavioral consequences of stress. To investigate this hypothesis, male Wistar rats were stressed (forced swimming, 15 min) and 24 h later received intra-BNST injections of vehicle, WB4101, RX821002, CGP20712, or ICI118,551, which are selective α1, α2, β1, and β2 adrenoceptor antagonists, respectively, 10 min before a 5-min forced swimming test. It was observed that administration of WB4101 (10 and 15 nmol), CGP20712 (5 and 10 nmol), or ICI118,551 (5 nmol) into the BNST reduced the immobility time of rats subjected to forced swimming test, indicating an antidepressant-like effect. These findings suggest that activation of α1, β1, and β2 adrenoceptors in the BNST could be involved in the development of the behavioral consequences of stress. © 2013 Wolters Kluwer Health | Lippincott Williams & Wilkins.

Rapid determination of furosemide by combined spot test/diffuse reflectance spectroscopy to detect doping in sport

This paper describes the development and application of a simple, cheap, and clean method for the quantification of furosemide in urine samples from athletes, to detect doping, using a combined spot test/diffuse reflectance spectroscopy procedure. The method is based on the complexation reaction of furosemide (5-(aminosulfonyl)-4-chloro-2-((furanylmethyl)amino)benzoic acid, dissolved in ethanol, with FeCl3 and the surfactant dodecyltrimethylammonium bromide (DTAB) in aqueous solution, yielding a colored compound on the surface of a filter paper. The reagent concentrations were optimized using a chemometric experimental design. The reflectometric measurements of the complex formed were carried out at 477nm. The linear range obtained was 1.65-9.00×10-3molL-1 of furosemide (R=0.997), and the detection and quantification limits were 4.9×10-4 and 1.62×10-3molL-1, respectively. The proposed method was successfully applied in the analysis of furosemide in spiked urine, demonstrating that it is a reliable alternative method for the detection of furosemide doping in sport. © 2012 Elsevier B.V..

Development and sensory analysis of shampoo for curly hair

Objective: This study aimed to compare the sensory performance of a shampoo formulation with Polyurethane-14, AMP-acrylates copolymer (PAAC) in relation to control formulation in curly and natural hair tresses. Methods: Curly and natural hair tresses (n = 8) of equal size and weight were pre-treated by washing with a standard shampoo. After the hair tresses were treated with a formulation containing polymer (formulation A) and compared to hair tresses treated with control formulation (Formulation B). Each panelist (n=2) is asked to indicate which tress performs better for each of seven sensory attributes evaluated (quantity and creamy foam, combing, wet touch, frizz formation, curl definition and volume). It was collected images of hair tresses at 0, 1, 2, 4 and 24 hours of washing, comparing the attributes: volume, frizz formation and curl definition. The results were analyzed using table to test of paired assessment, being: SUPERIOR results - 8 and 7 positive evaluations; SIMILAR results - 2 to 6 positive evaluations; INFERIOR results - 1 and 0 positive evaluations. Results: The addition of the PAAC on the shampoo formulation provided definition and modeling of curls, reducing volume and frizz in 24 hours. There was also lower foam formation in the formulation with polymer PAAC. However, it is important to note that this attribute has inversely proportional effect to the creamy foam, since more creamy foam, smaller quantity. Conclusions: It was concluded that the shampoo developed was effective in defining and modeling curl in natural and curly hair.

Development of equations to estimate microbial contamination in ruminal incubation residues of forage produced under tropical conditions using 15N as a label1

The objective of this study was to use 15N to label microbial cells to allow development of equations for estimating the microbial contamination in ruminal in situ incubation residues of forage produced under tropical conditions. A total of 24 tropical forages were ruminal incubated in 3 steers at 3 separate times. To determine microbial contamination of the incubated residues, ruminal bacteria were labeled with 15N by continuous intraruminal infusion 60 h before the first incubation and continued until the last day of incubation. Ruminal digesta was collected for the isolation of bacteria before the first infusion of 15N on adaptation period and after the infusion of 15N on collection period. To determine the microbial contamination of CP fractions, restricted models were compared with the full model using the model identity test. A value of the corrected fraction A was estimated from the corresponding noncorrected fraction by this equation: Corrected A fraction (ACPC) = 1.99286 + 0.98256 × A fraction without correction (ACPWC). The corrected fraction B was estimated from the corresponding noncorrected fraction and from CP, NDF, neutral detergent insoluble protein (NDIP), and indigestible NDF (iNDF) using the equation corrected B fraction (BCPC) = -17.2181 - 0.0344 × fraction B without correction (BCPWC) + 0.65433 × CP + 1.03787 × NDF + 2.66010 × NDIP - 0.85979 × iNDF. The corrected degradation rate of B fraction (kd)was estimated using the equation corrected degradation rate of B fraction (kdCPC) = 0.04667 + 0.35139 × degradation rate of B fraction without correction (kdCPWC) + 0.0020 × CP - 0.00055839 × NDF - 0.00336 × NDIP + 0.00075089 × iNDF. This equation was obtained to estimate the contamination using CP of the feeds: %C = 79.21 × (1 - e-0.0555t) × e-0.0874CP. It was concluded that A and B fractions and kd of CP could be highly biased by microbial CP contamination, and therefore these corrected values could be obtained mathematically, replacing the use of microbial markers. The percentage of contamination and the corrected apparent degradability of CP could be obtained from values of CP and time of incubation for each feed, which could reduce cost and labor involved when using 15N. © 2013 American Society of Animal Science. All rights reserved.

Mandibular growth and dentoalveolar development in the treatment of Class II, division 1, malocclusion using Balters Bionator according to the skeletal maturation

OBJECTIVE: The purpose of the study was to evaluate the influence of the skeletal maturation in the mandibular and dentoalveolar growth and development during the Class II, division 1, malocclusion correction with Balters bionator. METHODS: Three groups of children with Class II, division 1, malocclusion were evaluated. Two of them were treated for one year with the bionator of Balters appliance in different skeletal ages (Group 1: 6 children, 7 to 8 years old and Group 2: 10 children, 9 to 10 years old) and the other one was followed without treatment (Control Group: 7 children, 8 to 9 years old). Lateral 45 degree cephalometric radiographs were used for the evaluation of the mandibular growth and dentoalveolar development. Tantalum metallic implants were used as fixed and stable references for radiograph superimposition and data acquisition. Student's t test was used in the statistical analysis of the displacement of the points in the condyle, ramus, mandibular base and dental points. Analysis of variance one-fixed criteria was used to evaluate group differences (95% of level of significance). RESULTS: The intragroup evaluation showed that all groups present significant skeletal growth for all points analyzed (1.2 to 3.7 mm), but in an intergroup comparison, the increment of the mandibular growth in the condyle, ramus and mandibular base were not statically different. For the dentoalveolar modifications, the less mature children showed greater labial inclination of the lower incisors (1.86 mm) and the most mature children showed greater first permanent molar extrusion (4.8 mm).

Development and validation of a microbiological assay by turbidimetry to determine the potency of cefazolin sodium in the lyophilized powder form

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Development and validation of a rapid turbidimetric assay to determine the potency of ampicillin sodium in powder for injectable solution

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Neonatally induced mild diabetes: influence on development, behavior and reproductive function of female Wistar rats

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Development of a Methodology and Evaluation of Pesticides Against Aceria litchii and Its Predator Phytoseius intermedius (Acari: Eriophyidae, Phytoseiidae)

The litchi erineum mite, Aceria litchii (Keifer), is the major pest of litchi, Litchi chinensis Sonnerat (Sapindaceae). This study evaluated the effect of 11 pesticides on the survival of A. litchii as well as on the survival and reproduction of Phytoseius intermedius Evans& McFarlane, the predator most found in association with it in Brazil. The containment of A. litchii in small petri dishes whose bottoms were covered with a solidified paste made of a mixture of gypsum and activated charcoal (9: 1 in volume), kept humid, was shown to be adequate for this type of study. For the evaluation of the effect of pesticides on A. litchii, mites leaving the erinia from the pieces of litchi leaves (removed from the plants approximate to 24 h earlier) were sprayed under a Potter tower and immediately transferred to the 2.5-cm-diameter petri dishes. After 2, 12, 24, and 48 h of the application, the dishes were examined to evaluate the mite survival. The four pesticides causing the highest levels A. litchii mortality, as well as azadirachtin, were tested for the effect on P. intermedius. For this test, experimental units consisted of discs of uninfested litchi leaves also sprayed under a Potter tower before introducing the predators. Survival and oviposition of the predator were evaluated every 24 h for 5 consecutive days; viability of the eggs laid was also evaluated. Highest mortality of A. litchii occurred with the application of fenpyroximate, sulfur, abamectin, and hexythiazox. Azadirachtin was considered moderately harmful to the predator P. intermedius, whereas other pesticides were classified as harmful. Despite the low efficiency of azadirachtin in the control of the pest, its relative selectivity to P. intermedius would encourage the evaluation on field condition, especially for use in organic production systems.

Development of a CAD system for automatic classification of microcalcifications based on FPGA

This paper presents a Computer Aided Diagnosis (CAD) system that automatically classifies microcalcifications detected on digital mammograms into one of the five types proposed by Michele Le Gal, a classification scheme that allows radiologists to determine whether a breast tumor is malignant or not without the need for surgeries. The developed system uses a combination of wavelets and Artificial Neural Networks (ANN) and is executed on an Altera DE2-115 Development Kit, a kit containing a Field-Programmable Gate Array (FPGA) that allows the system to be smaller, cheaper and more energy efficient. Results have shown that the system was able to correctly classify 96.67% of test samples, which can be used as a second opinion by radiologists in breast cancer early diagnosis. (C) 2013 The Authors. Published by Elsevier B.V.