Determinação precoce do sexo fetal pela análise do DNA no plasma materno

Purpose: to verify the viability of early diagnosis of fetal gender in maternal plasma by the real-time polymerase chain reaction (real-time PCR) starting at the 5th week of pregnancy. Methods: peripheral blood was collected from pregnant women with single fetus starting at the 5th week of gestation. After centrifugation, 0.4 mL plasma was separated for fetal DNA extraction. The DNA was analyzed in duplicate by real-time PCR for two genomic regions, one of the Y chromosome and the other common to both sexes, through the TaqMan® method, which uses a pair of primers and a fluorescent probe. Patients who aborted were excluded. Results: a total of 79 determinations of fetal DNA in maternal plasma were performed in 52 pregnant women. The results of the determinations were compared to fetal gender after delivery. Accuracy according to gestational age was 92.6% (25 of 27 cases) at 5 weeks with 87% sensitivity, and 95.6% (22 of 23 cases) at 6 weeks with 92% sensitivity. Starting at the 7th week of pregnancy, accuracy was 100% (29 of 29 cases). Specificity was 100% regardless of gestational age. Conclusion: real-time PCR for the detection of fetal gender in maternal plasma starting at the 5th week of gestation has good sensitivity and excellent specificity. There was agreement of the results in 100% of the cases in which male gender was diagnosed, regardless of gestational age, and from the 7th week of gestation for female gender diagnosis.

Avoiding leukocyte contamination and early platelet activation in platelet-rich plasma.

The objective of this study was to describe a new platelet-rich plasma (PRP) protocol with a reduced concentration of leukocytes and intact platelets. We collected 8 mL of venous blood (VB) from marginal ear veins of 10 male New Zealand white rabbits in acid dextrose citrate Vacutainer tubes. Tubes were centrifuged at 302g for 10 minutes. All plasma was collected in plastic tubes to avoid buffy-coat contamination and centrifuged at 2862g for 5 minutes. A 10% calcium chloride activator (10 PRP:2 CaCl2) was added to the lower third of this plasma (PRP), and the PRP gel was obtained. Mean platelet count was 317.7 x 10(3) +/- 39.9/microL in VB and 1344.9 x 10(3) +/- 347.5/microL in PRP. Leukocyte counts were 3.96 x 10(3) +/- 2.01/microL and 0.46 x 10(3) +/- 0.45/microL in VB and PRP, respectively. Mean platelet enrichment was 327.4 +/- 97.8%. All differences were statistically significant (P > .05). This protocol is practical and reproducible, resulting in a high concentration of intact platelets to help tissue repair and low levels of leukocytes.

Acute phase proteins in canine lymphoma during antineoplastic chemotherapy

The lymphoma is the main hematopoietic tumor in dogs and it is characterized by the proliferation of cells from lymphoid tissue, histiocytes and its precursors. Animals with lymphoma often show changes in biochemical and hematological parameters such as non-regenerative normochromic normocytic anemia, hemolytic anemia, hypocalcaemia and monoclonal gammopathy. The development of tumor can cause alterations in serum concentrations of acute phase proteins (APPs), consequent of hepatocytes stimulus by cytokines of inflammatory action. This study aimed to quantify and qualify APPs in dogs with lymphoma, at diagnosis time and during the time of chemotherapy sessions. After syneresis, centrifugation and fractioning the serum samples of 10 healthy and 10 dogs with lymphomas, the proteins fractions were separated by polyacrilamide gel electrophoresis (SDS-PAGE) and its concentrations were determined by computer densitometry. Between 18 and 30 proteins were separated by eletrophoresis, with molecular weights ranging from 18 to 245 kDa (kilodaltons). The alpha-1-glicoprotein acid (AGP) and transferrin serum concentration showed significantly higher in dogs with lymphoma, when compared with healthy dogs at diagnosis. The alpha-1-antitripsin (AAT) serum concentrations showed significantly higher in healthy dogs, when compared with dogs with lymphoma at diagnosis. The dogs with lymphoma the albumin did not appear as negative APP. On the other hand, transferrin appeared as positive AAP at diagnosis time and during the chemotherapy sessions. Healthy dogs had AAT serum concentrations significantly higher when compared to dogs with lymphoma at diagnosis. So, in this trial, it is suggested that this protein has been shown as a negative APP in the dogs with lymphoma. These dogs presented significantly higher AGP serum concentrations, in relation to healthy dogs at diagnosis, evidencing this protein APP positive behavior in neoplasm.

Embryos refrozen-thawed by vitrification lead to live births: Case report

Objective: To describe two successful cases of utilizing refrozen blastocysts by vitrification derived from supernumerary embryos. Design: Case report. Setting: Private fertility clinic. Subjects: Two infertility couple. Interventions: Refrozen blastocysts by vitrification derived from supernumerary embryos. Main outcome measures: Obstetric and pediatric results. Results: Two pregnancies obtained from refrozen-warmed blastocysts led to two healthy babies being born without clinical or genetic problems. Conclusion: These case reports support the notion of safely repeating cryopreservation. However, despite these favorable results, there is still a need for prospective controlled studies on the obstetric and neonatal repercussions of refreezing and of vitrification in particular. © 2010 Middle East Fertility Society.

Um novo método laboratorial para diagnóstico de parasitos gastrointestinais em cães

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

Pharmacokinetics of dexmedetomidine administered intravenously in isofluraneanesthetized cats

Objective-To determine the pharmacokinetics of dexmedetomidine administered as a short-duration IV infusion in isoflurane-anesthetized cats. Animals-6 healthy adult domestic female cats. Procedures-Dexmedetomidine hydrochloride was injected IV (10 μg/kg over 5 minutes [rate, 2 μg/kg/min]) in isoflurane-anesthetized cats. Blood samples were obtained immediately prior to and at 1, 2, 5, 6, 7, 10, 15, 30, 60, 90, 120, 240, and 480 minutes following the start of the IV infusion. Collected blood samples were transferred to tubes containing EDTA, immediately placed on ice, and then centrifuged at 3,901 X g for 10 minutes at 4°C. The plasma was harvested and stored at -20°C until analyzed. Plasma dexmedetomidine concentrations were determined by means of liquid chromatography-mass spectrometry. Dexmedetomidine plasma concentration-time data were fitted to compartmental models. Results-A 2-compartment model with input in and elimination from the central compartment best described the disposition of dexmedetomidine administered via short-duration IV infusion in isoflurane-anesthetized cats. Weighted mean ± SEM apparent volume of distribution of the central compartment and apparent volume of distribution at steady-state were 402 ± 47 mL/kg and 1,701 ± 200 mL/kg, respectively; clearance and terminal half-life (harmonic mean ± jackknife pseudo-SD) were 6.3 ± 2.8 mL/min/kg and 198 ± 75 minutes, respectively. The area under the plasma concentration curve and maximal plasma concentration were 1,061 ± 292 min·ng/mL and 17.6 ± 1.8 ng/mL, respectively. Conclusions and Clinical Relevance-Disposition of dexmedetomidine administered via short-duration IV infusion in isoflurane-anesthetized cats was characterized by a moderate clearance and a long terminal half-life.

Anti-platelet activity of infusion of Tithonia diversifolia's leaves

Tithonia diversifolia, also known as Mexican arnica, has been used in traditional medicine to treat inflammatory refractory with absence of citotoxicity. The possible health risks associated with the consumption of ingestion of the infusion (tea) plant makes it is necessary to identify the potential pharmacological activity or toxicity to prove certain plants that are acclimated in Brazil. Considering the limited number of pharmacological studies regarding the Tithonia diversifolia, the aim of this study was evaluate the effects of this infusion in platelet aggregation. Venous blood was collected with informed consent from healthy volunteers who denied taking any medication in the previous 14 days. Whole blood was transferred into polypropylene tubes containing one-tenth of final volume of acid citrate dextrose (ACD-C; citric acid 3%, trisodium citrate 4%, glucose 2%; 1:9 v/v) and centrifuged at 200g for 15 min. Platelet rich plasma was added of wash buffer solution (NaCl 140mM, KCl 5mM, sodium citrate 12mM, glucose 10mM and saccharose 12mM; pH 6; 5:7 v/v) and centrifuged at 800g for 12 min at 20°C. Platelet pellet was gently resuspended in Krebs-Ringer solution and counts were performed on a Neubauer chamber. Aggregation assay was carried out with 400 μL of platelet suspension (1.2x10 8 platelets/mL) in a cuvette at 37°C with constant stirring. Platelet suspension was incubated for 3 min with aqueous extract infusion (0.6-20μg/mL) prior to addition of thrombin (100 mU/mL). Percentage of platelet aggregation was recorded with an aggregometer (Chrono-log Lumi-Aggregometer model 560-Ca, USA). Our results show an inhibition of thrombin induced platelet aggregation in the presence of 0.6-20 ug/mL Tithonia diversifolia infusion leaves. The Tithonia diversifolia infusion leaves inhibits thrombin induced washed platelet aggregation.

Effect of Removing Seminal Plasma Using a Sperm Filter on the Viability of Refrigerated Stallion Semen

Cooling of equine semen obtained from some stallions results in lower seminal quality and viability when the seminal plasma (SP) is present. The objective of this study was to evaluate the effect of the removal of SP using a Sperm Filter on the viability of cooled stallion semen. For this purpose, 31 stallions were used. Their ejaculates were divided into three groups: CN, semen was diluted with an extender; FLT, SP was removed by filtration; and CT, SP was removed by centrifugation and cooled to 15°C for 24 hours. Sperm kinetics and plasma membrane integrity were evaluated immediately after collection (T0) and after 24 hours of refrigeration (T1). No difference (P > .05) was noted at T1 for total sperm motility (TM), progressive sperm motility, or plasma membrane integrity when semen samples from all the stallions were analyzed. However, when samples from stallions termed bad coolers were analyzed (TM = <30% at T1), a difference was observed in TM and progressive sperm motility for CN compared with FLT and CT at T1. Sperm recovery was greater when SP was removed using the filter (FLT) to that when the SP was removed by centrifugation (CN) (89% vs. 81%). Thus, we concluded that filtering with a Sperm Filter is an efficient and practical method for removal of SP from stallion ejaculates, with lower sperm loss than centrifugation. We also found that the presence of SP reduces the quality and viability of cooled semen from stallions whose semen is sensitive to the process of refrigeration. © 2013 Elsevier Inc.

Lack of reliability of nanotechnology in the of free plasma DNA in samples of patients with prostate cancer

Background: Several studies seek biological markers that give diagnostic and degree of tumor development. The aim of this study was to validate the determination of plasma DNA using nanotechnology (Nanovue™-NV) in samples of 80 patients with prostate cancer. Methods. Blood samples of 80 patients of the Urology Ambulatory of Faculdade de Medicina do ABC with prostate cancer confirmed by anatomical-pathology criteria were analyzed. DNA extraction was performed using a GFX TM kit (Amersham Pharmacia Biotech, Inc, USA) following the adapted protocol. Plasma was subjected to centrifugation. Results: There was a big difference between the first and the second value obtained by NanoVue Only two samples had no differences between duplicates. Maximum difference between duplicates was 38 μg/mL. Average variation between 51 samples was 10.29 μg/mL, although 21 samples had differences above this average. No correlation was observed between pDNA obtained by traditional spectrophotometry and by nanotechnology. Conclusion: Determination of plasma DNA by nanotechnology was not reproducible. © 2013 Moreno et al; licensee BioMed Central Ltd.

Nebular gas drag and planetary accretion with eccentric high-mass planets

Aims.We investigate the dynamics of pebbles immersed in a gas disk interacting with a planet on an eccentric orbit. The model has a prescribed gap in the disk around the location of the planetary orbit, as is expected for a giant planet with a mass in the range of 0.1-1 Jupiter masses. The pebbles with sizes in the range of 1 cm to 3 m are placed in a ring outside of the giant planet orbit at distances between 10 and 30 planetary Hill radii. The process of the accumulation of pebbles closer to the gap edge, its possible implication for the planetary accretion, and the importance of the mass and the eccentricity of the planet in this process are the motivations behind the present contribution. Methods. We used the Bulirsch-Stoer numerical algorithm, which is computationally consistent for close approaches, to integrate the Newtonian equations of the planar (2D), elliptical restricted three-body problem. The angular velocity of the gas disk was determined by the appropriate balance between the gravity, centrifugal, and pressure forces, such that it is sub-Keplerian in regions with a negative radial pressure gradient and super-Keplerian where the radial pressure gradient is positive. Results. The results show that there are no trappings in the 1:1 resonance around the L 4 and L5 Lagrangian points for very low planetary eccentricities (e2 < 0.07). The trappings in exterior resonances, in the majority of cases, are because the angular velocity of the disk is super-Keplerian in the gap disk outside of the planetary orbit and because the inward drift is stopped. Furthermore, the semi-major axis location of such trappings depends on the gas pressure profile of the gap (depth) and is a = 1.2 for a planet of 1 MJ. A planet on an eccentric orbit interacts with the pebble layer formed by these resonances. Collisions occur and become important for planetary eccentricity near the present value of Jupiter (e 2 = 0.05). The maximum rate of the collisions onto a planet of 0.1 MJ occurs when the pebble size is 37.5 cm ≤ s < 75 cm; for a planet with the mass of Jupiter, it is15 cm ≤ s < 30 cm. The accretion stops when the pebble size is less than 2 cm and the gas drag dominates the motion. © 2013 ESO.

New seminal plasma removal method for freezing stallion semen

Seminal plasma removal, an indispensable step in equine semen cryopreservation, is usually done by centrifugation, but this might cause mechanical damage to sperm. A new method for seminal plasma removal from stallion semen, namely a filter composed of a synthetic hydrophilic membrane (Sperm Filter, BotuPharma, Botucatu, Sao Paulo, Brazil), was recently proposed. The objective of this study was to test the use of the Sperm Filter in the removal of seminal plasma before freezing stallion semen. Ejaculates from 31 stallions were divided into two groups and cryopreserved. In group 1 (G1), seminal plasma was removed with the Sperm Filter, and in group 2 (G2), seminal plasma was removed by centrifugation (600× g for 10 minutes). There were no differences (P < 0.05) between G1 and G2 in sperm kinetic parameters or plasma membrane integrity before or after cryopreservation. However, sperm recovery rate was higher (P < 0.05) for G1 versus G2 (mean ± SD, 89.4 ± 7.4% vs. 80.9 ± 5.5%). Therefore, the Sperm Filter was as efficient as centrifugation in removing seminal plasma from the stallion ejaculate. However, filtering was more practical and had significantly fewer sperm lost than the centrifugation technique. © 2013 Elsevier Inc.

Occurrence of Cu and Cr in the sedimentary humic substances and pore water from a typical sugar cane cultivation area in São Paulo, Brazil

Purpose: The purpose of this paper is to study the interactions of sedimentary humic substances (SHS) from a sugarcane cultivation area with Cu(II) and Cr(III) and to evaluate the occurrence of these metals in the pore water and SHS. Materials and methods: For this study, the northwestern region of the State of São Paulo, Brazil, which is considered the region with the highest production of sugar cane in the state, was selected. Samples of sediment were collected from four sampling sites in the Preto, Turvo, and Grande rivers. The SHS and pore water were extracted from the sediment using the method suggested by the International Humic Substances Society and centrifugation, respectively. The complexing capacity (CC) of the SHS for Cu(II) and Cr(III) was determined by individually titrating these metals with an ultrafiltration system using tangential flow. The total concentrations of Cr and Cu were determined for the pore water, sediments, and humic substances with graphite furnace atomic absorption spectrometry and Zeeman background correction after an acid digestion, according to the methods described in US EPA Method 3050B. Results and discussion: The SHS from a site in the Turvo River, which is typically cultivated with sugarcane, possessed the highest concentration of Cu bound to SHS (25.0%), the largest CC (0.63 mmol Cu g-1 HS) and the highest concentration of this metal in the pore water (1.38 mg Cu Kg-1 sed.). For Cr, the SHS collected from a location on the Preto River dam had the largest CC (0.90 mmol Cr g-1 HS) and the lowest Cr content in the pore water (0.29 mg Cr Kg-1 sed.), indicating that there was an inverse relationship between the CC and the concentration of metal available in the pore water. Conclusions: Sedimentary humic substances might be one of the regulatory factors controlling the availability of Cu and Cr in the sediments found in a typical region that has been planted with sugarcane. Distinct behaviors were observed between the two elements investigated; higher CC and a larger fraction of Cu(II) were found in the pore water of samples originating from sugarcane crops. The opposite behavior was observed for the Cr(III) species. © 2013 Springer-Verlag Berlin Heidelberg.

Monitoramento da qualidade de hemocomponentes produzidos no Hemocecentro da FMB - UNESP

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

Preparação e caracterização de óxido de zinco nanoestruturado

Pós-graduação em Ciência e Tecnologia de Materiais - FC

Desenvolvimento e caracterização físico-química de sistemas líquido cristalinos contendo palmitato de retinol: quantificação química, avaliação da atividade antioxidante, estudos de liberação e bioadesão in vitro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)