Development of a real-time PCR method for rapid sexing of human preimplantation embryos

Genes on the X chromosome are known to be responsible for more than 200 hereditary diseases. After IVF, the simple selection of embryo sex before uterine transfer can prevent the occurrence of affected offspring among couples at risk for these genetic disorders. The aim of this investigation was to develop a rapid method of preimplantation genetic diagnosis (PGD) using real-time polymerase chain reaction (PCR) for the sexing of human embryos, and to compare it to the fluorescence in-situ hybridization technique, considered to be the gold standard. After biopsies were obtained from 40 surplus non-viable embryos for transfer, a total of 98 blastomeres were analysed. It was possible to analyse 24 embryos (60%) by both techniques, generating a total of 70 blastomeres (35 per technique), white 28 blastomeres from 16 embryos (40%) were analysed only by real-time PCR. A rapid and safe method was developed in the present study for the sexual diagnosis of a single human cell (blastomere and buccal cell) using the emerging technology of real-time PCR. (C) 2009, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Is urine interleukin-8 level a reliable laboratory test for diagnosing late onset sepsis in premature infants?

The present study is aimed to determine serum and urine interleukin-8 (IL-8) levels in premature infants with late onset sepsis (LOS) and to evaluate if urine IL-8 is a useful test for LOS diagnosis. Fifty-six premature infants admitted to the NICU over 1 year had serum and urine IL-8 determined by ELISA. They were divided into three groups: I definite sepsis, II probable sepsis and III non-infected. Results were expressed as mean or median. Differences between groups were assessed by ANOVA, Kruskal-Wallis ANOVA and Dunns Method. Sensitivity, specificity and positive and negative predictive values were calculated and a receiver operator characteristic curve was constructed to determine serum and urine IL-8 accuracy. There were no differences between groups for birth weight, and gestational and post-natal age. Median serum and urine IL-8 levels were significantly higher in GI and GII: 929 x 906 x 625pg/ml; P=0.024, and 249 x 189 x 42pg/mgCr; P< 0.001. Optimal cut-off point was 625pg/ml for serum IL-8 with 69 sensitivity and 75pg/mgCr for urine IL-8 with 92 sensitivity. IL-8 can be determined in urine from premature infants with LOS and is an accurate and feasible diagnosis method.

Indigestible cellulose and lignin in determining feces production and apparent digestibility in horses

Foram utilizados quatro cavalos castrados sem raça definida, distribuídos em blocos casualizados. Objetivou-se estudar a viabilização dos indicadores internos, celulose (CELi) e lignina indigestíveis (LIGi), para predizer a digestibilidade em cavalos. Os tratamentos consistiram na determinação da digestibilidade por método direto com a coleta total de fezes (CT) e indireto pelo uso CELi e LIGi obtidos pelas técnicas in situ (IS) na cavidade ruminal de bovinos e in vivo (IV) nos equinos por meio do saco de náilon móvel (SNM). A produção fecal e taxa de recuperação (p > 0,05) médios encontrados foram de 2,80 kg na MS e 101%, respectivamente. As estimativas dos CD dos nutrientes (p > 0,05) foram adequadamente preditos pela CELi e LIGi, obtidos in situ e in vivo, no qual os valores médios observados foram de 52,63, 54,17, 64,90, 43,73 e 98,28% para MS, MO, PB, FDN e Amido, respectivamente. Concluiu-se que a CELi e LIGi podem ser obtidas in vivo por meio do SNM em equinos, para predizer os coeficientes de digestibilidade de nutrientes, consumindo dieta mista.

Histological Characterization and Immunohistochemical Localization of Ki-67 and VEGF Factors in Bitches' Corpus Luteum at Cyclic and Early and Late Gestational Diestrous

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

Relative expression of insulin like growth factor I (IGFI) and follicle stimulating hormone receptor (FSHR) in follicles and ovarian tissue from Bos primigenius indicus (Nelore)

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

Bone healing in critical-size defects treated with either bone graft, membrane, or a combination of both materials: a histological and histometric study in rat tibiae

Objectives: The aim of the present investigation was to histologically analyze the effect of using lyophilized bovine bone (GenOx (R) organic matrix) with (or without) guided tissue regeneration (using a decalcified cortical osseous membrane [GenDerm (R)]) on bone healing in surgically created critical-size defects created in rat tibia.Material and methods: Surgical critical-size bone defects were created in 64 animals that were randomly divided into four groups: group I (control); group II (defect filled with GenOx (R)); group III (defect covered by GenDerm (R)); group IV (defect filled with GenOx (R) and covered by GenDerm (R)). Animals were killed at 30 or 90 days post-surgery. The specimens were embedded in paraffin, serially cut, and stained with hematoxylin and eosin for analysis under light microscopy. The formation of new bone in the cortical area of the defect was histomorphometrically evaluated.Results: All experimental groups demonstrated superior bone healing compared with the control group. However, group IV samples showed evidence of more advanced healing at both 30 and 90 days post-surgery as compared with the other experimental groups.Conclusions: The bovine organic bone graft GenOx (R) associated with GenDerm (R) this produced the best treatment results in the case of critical-size defects in rat tibia.

In vitro comparison of laser fluorescence performance with visual examination for detection of occlusal caries in permanent and primary molars

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evaluation of marginal microleakage in Class II cavities: Effect of microhylolrid, flowable, and compactable resins

Objective: the goal of the present study was to evaluate the microleakage on the cementum/dentin and enamel surfaces in Class 11 restorations, using different kinds of resin composite (microhybrid, flowable, and compactable). Method and materials: Forty human caries-free molars were extracted and selected. Eighty Class 11 standardized cavities were made in the cervical wall at the cementoenamel junction (CEJ) and at the mesial and distal surfaces. The teeth were divided into four groups: G1 - adhesive system + microhybrid resin composite Z100; G2 - adhesive system + compactable resin composite Prodigy Condensable; G3 - adhesive system + flowable resin composite Revolution + Z1 00 resin composite; G4 - adhesive system + Revolution fluid resin + compactable resin composite Prodigy Condensable. The adhesive system used in this study was Scotchbond Multi-Purpose Plus. The specimens were thermocycled in baths of 5degreesC and 55degreesC for 1,000 cycles and immersed in 50% silver nitrate solution. The specimens then were sectioned and evaluated on degree of dye penetration. Results: the results were evaluated using the nonparametric Kruskall-Wallis test, which showed a statistically significant difference between groups G1 and G4, G2 and G4, and G3 and G4. Conclusions: None of the materials was able to eliminate the marginal microleakage at the cervical wall; the application of a low-viscosity resin composite combined with a compactable resin composite significantly decreased the microleakage.

Ability of Gutta-Percha and Resilon to Fill Simulated Lateral Canals by Using the Obtura II System

Introduction: The aim of this study was to evaluate the ability of Resilon (Resilon Research, LLC, North Branford, CT) and 2 types of gutta-percha to fill simulated lateral canals when using the Obtura II system (Model 823-700; Obtura Spartan, Fenton, MO). Methods: Forty-five human single-rooted teeth were selected and subjected to root canal preparation. After that, simulated lateral canals were made at 2, 5, and 8 mm from the working length (WL). The specimens were divided into 3 groups (n = 15) according to the filling material used: Obtura Flow 150 gutta-percha (Obtura flow), Odous Endo Flow gutta-percha (Odous; Odous de Deus Ind e Corn. Ltda Belo Horizonte, MG, Brazil), and Resilon pellets (Resilon). Root canals were filled using the Obtura II system with the tip inserted to 3 mm from the WL. No sealer was used for root canal obturation. Specimens were subjected to a tooth decalcification and clearing method, and filling of the lateral canals was analyzed by digital radiography and photographs. The measurement of lateral canal filling was done using Image Tool software (UTHSCSA Image Tool for Windows version 3.0, San Antonio, TX). Data were statistically analyzed with the Kruskal-Wallis test at 5% significance. Results: All materials showed an ability to penetrate into the simulated lateral canals, with a minimum percentage of 73% in all thirds of the root canal. Conclusions: It was concluded that gutta-percha and Resilon are solid core materials with a lateral canal filling ability when used with the Obtura II system. (J Endod 2012;38:676-679)

Customized fiber glass posts. Fatigue and fracture resistance

Purpose: To evaluate the root fracture strength of human single-rooted premolars restored with customized fiberglass post-core systems after fatigue simulation. Methods: 40 human premolars had their crowns cut and the root length was standardized to 13 mm. The teeth were endodontically treated and embedded in acrylic resin. The specimens were distributed into four groups (n=10) according to the restorative material used: prefabricated fiber post (PFP), PFP+accessory fiber posts (PFPa), PFP+unidirectional fiberglass (PFPf), and unidirectional fiberglass customized post (CP). All posts were luted using resin cement and the cores were built up with a resin composite. The samples were stored for 24 hours at 37 degrees C and 100% relative humidity and then submitted to mechanical cycling. The specimens were then compressive-loaded in a universal testing machine at a crosshead speed of 0.5 mm/minute until fracture. The failure patterns were analyzed and classified. Data was submitted to one-way ANOVA and Tukey's test (alpha= 0.05). Results: The mean values of maximum load (N) were: PFP - 811.4 +/- 124.3; PFPa - 729.2 +/- 157.2; PFPf - 747.5 +/- 204.7; CP - 762.4 +/- 110. Statistical differences were not observed among the groups. All groups showed favorable restorable failures. Fiberglass customized post did not show improved fracture resistance or differences in failure patterns when compared to prefabricated glass fiber posts. (Am J Dent 2012;25:35-38).

GFAAS Determination of Zinc in Fish Feed and Feces Using Slurry Sampling

This paper presents a simple, fast, and sensitive method to determine zinc in samples of feces and fish feed by electrothermal atomic absorption spectrometry through the direct introduction of slurries of the samples into the spectrometer's graphite tube. The procedure is based on the injection of 10 mu L of an acidified aqueous solution containing 0.50% w/v of feces or feed and 0.50% v/v HNO(3) into graphite tube. The limits of detection and quantification calculated for 20 readings of the blank of the standard slurries (0.50% w/v of feces or feed devoid of zinc) were 0.04 and 0.13 mu g L(-1) for the standard feces slurries and 0.05 and 0.17 mu g L(-1) for the standard feed slurries. The proposed method was applied in studies of digestibility of zinc in different fish feeds, and their results proved compatible with that obtained from samples mineralized by acid digestion using microwave oven.

Purification and characterization of the exopolygalacturonase produced by Aspergillus giganteus in submerged cultures

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

Microhabitat selection and daily movements of two rodents (Necromys lasiurus and Oryzomys scotti) in Brazilian Cerrado, as revealed by a spool-and-line device

Studies on patterns of habitat use by mammals are necessary for understanding the mechanisms involved in their distribution and abundance. In this study, we used the spool-and-line method to investigate habitat utilization by two sigmodontine rodents from Brazilian Cerrado, Necromys lasiurus and Oryzomys scotti. We conducted the study in a Cerrado area in central Brazil (15 degrees 56'S and e 47 degrees 56'W) where the animals were caught in an area of 7.68 ha of Cerrado sensu stricto. Captured individuals were marked, equipped with a spool-and-Line device, and released at the same capture point. The next day we followed the thread to record their daily movements and find their nests. To investigate microhabitat selection we compared habitat characteristics along traits of each studied species with general habitat characteristics of the study area. Although the mean 24-h distance was greater for N. lasiurus (mean +/- SE: 41.9 +/- 42.2 m, N=3) than for O. scotti (28.7 +/- 14.2 m, N=6) this difference was not significant (Mann-Whitney test, U=26, P>0.6). We detected significant differences among observed microhabitats variables of both species and available microhabitat characteristics as determined by discriminant analysis (Wilks's lambda F=3.001; df=14, 116; P<0.001). Both species were associated to microhabitat characteristics whose values differed markedly from the overall available habitat. Along the first canonical discriminant function of the DFA both them were associated with greater grass height than the mean height available and along the second axis N. lasiurus selected areas with higher fruit availability and more shelters than those selected by 0. scotti. For stronger inferences regarding differential patterns of habitat utilization by Cerrado rodents we suggest the simultaneous use of both spool-and-line and standard trapping methods. (c) 2005 Deutsche Geseltschaft fur Saugetierkunde. Published by Elsevier GmbH. ALL rights reserved.

Simultaneous Determination of Phenanthrene and Benzo(a)pyrene in Water Samples by Synchronous Fluorescence Spectroscopy

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

Dentine microhardness after different methods for detection and removal of carious dentine tissue

There are several methods for identifying carious dentinal tissue aiming to avoid removal of healthy dentinal tissue. Objectives: The purpose of this study was to test different methods for the detection of carious dentinal tissue regarding the amount of carious tissue removed and the remaining dentin microhardness after caries removal. Material and methods: The dentin surfaces of 20 bovine teeth were exposed and half of the surface was protected with nail polish. Cariogenic challenge was performed by immersion in a demineralizing solution for 14 days. After transverse cross-section of the crown, the specimens were divided into four groups (n=10), according to the method used to identify and remove the carious tissue: "Papacarie", Caries-detector dye, DIAGNOdent and Tactile method. After caries removal, the cross-sectional surface was included in acrylic resin and polished. In a microhardness tester, the removed dentin thickness and the Vickers microhardness of the following regions were evaluated: remaining dentin after caries removal and superficial and deep healthy dentin. Results: ANOVA and Tukey's test (alpha=0.05) were performed, except for DIAGNOdent, which did not detect the presence of caries. Results for removed dentin thickness were: "Papacarie" (424.7 +/- 105.0; a), Caries-detector dye (370.5 +/- 78.3; ab), Tactile method (322.8 +/- 51.5; bc). Results for the remaining dentin microhardness were: "Papacarie" (42.2 +/- 10.5; bc), Caries-detector dye (44.6 +/- 11.8; bc), Tactile method (24.3 +/- 9.0; d). Conclusions: DIAGNOdent did not detect the presence of carious tissue; Tactile method and "Papacarie" resulted in the least and the most dentinal thickness removal, respectively; Tactile method differed significantly from "Papacarie" and Caries-detector dye in terms of the remaining dentin microhardness, and Tactile method was the one which presented the lowest microhardness values.